TWI825021B - Mmp13 binding immunoglobulins - Google Patents

Abstract

The present invention relates to immunoglobulins that specifically bind MMP13 and more in particular to polypeptides, nucleic acids encoding such polypeptides; to methods for preparing such polypeptides; to compositions and in particular to pharmaceutical compositions that comprise such polypeptides, for prophylactic, therapeutic or diagnostic purposes. In particular, the immunoglobulins of the present invention inhibit an activity of MMP13 and preferably are also stable.

Description

Immunoglobulin that binds to MMP13

The present invention relates to immunoglobulins that bind to MMP13, and more particularly to polypeptides that comprise or consist essentially of one or more such immunoglobulins (also referred to herein as "immunoglobulins of the invention" and "the immunoglobulins of the invention", respectively). Composed of the polypeptide of the invention"). The invention also relates to constructs comprising such immunoglobulins or polypeptides and nucleic acids encoding such immunoglobulins or polypeptides (also referred to herein as "nucleic acids of the invention"); preparation of such immunoglobulins, polypeptides and constructs methods; host cells expressing or capable of expressing such immunoglobulins or polypeptides; compositions and particularly pharmaceutical compositions including such immunoglobulins, polypeptides, constructs, nucleic acids and/or host cells; and immunoglobulins Use of proteins, polypeptides, constructs, nucleic acids, host cells and/or compositions, particularly for prophylactic and/or therapeutic purposes, such as those mentioned herein. Other aspects, implementation aspects, advantages and applications of the present invention will become apparent from the further description herein.

Osteoarthritis (OA) is one of the most common causes of disability worldwide. It affects 30 million Americans and is the most common joint disorder. It is expected to affect more than 20% of the US population by 2025. The disease is not systemic and is usually localized to a few joints. However, the disease can occur in all joints, most commonly the knees, hips, hands, and spine. OA is characterized by the progressive erosion of articular cartilage (the cartilage covering bones), leading to chronic pain and disability. Eventually, the disease leads to complete destruction of the articular cartilage, hardening of the underlying tissue, and formation of osteophytes, all of which lead to loss of motion and pain. Pain is the most obvious symptom of OA and the most common reason patients seek medical help. There is no cure for OA; disease management is limited to the best palliative care and rarely addresses the underlying causes of disease progression. Disease-modifying anti-osteoarthritis drugs (DMOADs), which can be defined as drugs that inhibit the progression of structural disease and also ideally improve symptoms and/or function, are gaining popularity. DMOADs are likely to be prescribed long-term in this chronic condition in an aging population, thus requiring excellent safety data in a population with multiple comorbidities and potential for drug-drug interactions. Osteoarthritis can be defined as a diverse group of symptoms characterized by a combination of joint symptoms, signs arising from defects in articular cartilage, and changes in adjacent tissues, including bone, tendons, and muscles. The largest components in articular cartilage are proteoglycans and collagen (the most important is collagen II). The main proteoglycan in cartilage is aggrecan. Although disease initiation can be multifactorial, cartilage destruction appears to be the result of uncontrolled proteolytic extracellular matrix destruction (ECM). As mentioned above, the main component of the cartilage extracellular matrix is aggrecan (Kiani et al. 2002 Cell Research 12:19-32). This molecule is important in the normal function of articular cartilage because it provides a hydrated gel structure that gives the cartilage its load-bearing properties. Aggrecan is a large multimodular molecule (2317 amino acids) expressed in chondrocytes. Its core protein is composed of three globular domains (G1, G2 and G3) and a large extended region between G2 and G3 for glycosaminoglycan chain attachment. This extension contains two domains, one substituted by a keratan sulfate chain (KS domain) and one substituted by a chondroitin sulfate chain (CS domain). The CS domain has 100 to 150 glycosaminoglycan (GAG) chains attached to it. Aggrecan forms a large complex with hyaluronic acid, in which 50 to 100 aggrecan molecules interact through the G1 domain and link the protein with one hyaluronic acid molecule. Upon ingestion of water (due to the GAG content), these complexes form a reversibly deformable gel that resists compression. The structure, fluid retention and function of articular cartilage are related to the matrix content of aggrecan and the amount of chondroitin sulfate bound to the intact core protein. Type II collagen (collagen II, Col II) accounts for 50% of articular cartilage. Collagen fibrils form a network that allows cartilage to trap proteoglycans and provide tissue strength. Collagen is a structural protein composed of three right-handed bundles of parallel left-handed polyproline type II (PPII) helices. Since the PPII helices are tightly packed within the triple helix, every third residue (which is an amino acid) is Gly (glycine). Because glycine is the smallest amino acid without side chains, it plays a unique role in fibrous structural proteins. In collagen, Gly must be in every third position because the assembly of the triple helix places this residue in the interior (on the axis) of the helix, where there is no room for more than the single hydrogen atom of glycine. Large side groups. OA is characterized by 1) degradation of aggrecan, progressive release of domains G3 and G2 (resulting in "debulking" of cartilage) and eventual release of domain G1, and 2) degradation of collagen, irreversible destruction of cartilage structure. There is compelling evidence that matrix metalloproteinases (MMPs) play an important role in the tissue destruction associated with OA. MMPs are a family of zinc-dependent endopeptidases involved in extracellular matrix degradation and tissue remodeling. There are approximately 28 MMP family members, which can be classified into various subgroups, including collagenases, gelatinases, stromelysins, membrane-type MMPs, stromelysins, enamelysins, and others. Collagenase including MMP1, MMP8, MMP13 and MMP18 can degrade triple helical fibrillar collagen into special 3/4 and 1/4 fragments. In addition, MMP14 also showed evidence of cleaving fibrillar collagen and MMP2 was also able to dissolve collagen fibers. MMPs have long been considered as attractive therapeutic targets for the treatment of OA. However, broad-spectrum MMP inhibitors developed to treat arthritis failed in clinical trials due to painful joint stiffness side effects known as musculoskeletal syndrome (MSS). MSS is believed to be caused by non-selective inhibition of various MMPs. Nam et al. (2017 Proc Natl Acad Sci USA 113:14970-14975) describe nanobodies apparently specifically targeting the active site of MMP14. Therapeutic intervention of OA is also hindered by the difficulty of targeting articular cartilage with drugs. Because articular cartilage is an avascular and alymphoid tissue, traditional drug delivery routes (oral, intravenous, intramuscular) ultimately rely on transsynovial transfer of drugs via passive diffusion from synovial capillaries to cartilage. Therefore, in the absence of a mechanism to selectively target drugs to cartilage, systemic exposure of the body to high drug concentrations is necessary to achieve sustained intra-articular therapeutic doses. Most traditional therapies used for OA suffer from severe toxicity due to high systemic exposure. Additionally, most newly developed DMOADs have short residence times in joints, even when administered intra-articularly (Edwards 2011 Vet. J. 190:15-21; Larsen et al. 2008 J Pham Sci 97:4622 -4654). Intra-articular (IA) delivery of therapeutic proteins is limited by their rapid clearance from the joint space and lack of retention within cartilage. The synovial residence time of drugs in joints is often less than 24 hours. Due to the rapid clearance of most IA injected drugs, frequent injections are always required to maintain effective concentrations (Owen et al. 1994 Br. J. Clin Pharmacol. 38:349-355). However, frequent IA injections are undesirable due to pain and discomfort, and injections may pose challenges to patient compliance and introduce the risk of joint infection. There is still a need for effective DMOAD.

It is an object of the present invention to provide polypeptides against OA which have improved preventive, therapeutic and/or pharmacological properties compared to prior art amino acid sequences and antibodies, as well as other advantageous properties such as, for example, improved ease of preparation. , good stability and/or reduced cost of goods). In particular, it is an object of the present invention to provide immunoglobulin single variable domains (ISVDs) and polypeptides comprising them for the inhibition of MMPs and in particular for the inhibition of MMP13. The inventors speculate that the best area to inhibit the enzymatic activity of MMP13 would be to increase ISVD against the catalytic pocket. However, this becomes a major challenge. In particular, MMP13 is secreted as an inactive pro-form (proMMP13), in which the prodomain obscures the catalytic pocket and, as such, makes the catalytic pocket less accessible to enhance immune responses. On the other hand, activated MMP13 has a short half-life, mainly due to autologous proteolysis. Furthermore, even after the inventors overcame the first two problems, the high sequence conservation of the catalytic domain between various species resulted in foregoing a robust immune response. Finally, the inventors were able to address these challenges through innovatively developed new tools and unconventional screening methods. ISVDs were isolated from different screening campaigns and further engineered to possess diverse and advantageous characteristics, including stability, affinity and inhibitory activity. Monovalent ISVDs of the invention that bind to MMP13 outperform comparative drugs. Biparatopic peptides containing ISVDs less suitable for inhibiting MMP13 activity were even more potent. Therefore, the present invention relates to a polypeptide comprising at least one immunoglobulin single variable domain (ISVD) that binds to a matrix metalloproteinase (MMP) and preferably to the matrix metalloproteinase MMP13. The invention also includes polypeptides comprising two or more ISVDs that each individually specifically bind to MMP13, wherein a) at least the "first" ISVD is bound to a first epitope, epitope, part, domain, Subunit or conformation specific binding; and wherein b) at least the "second" ISVD specifically binds to a second epitope, epitopic region, portion, domain, subunit or conformation of MMP13 that is different from A first epitope, epitope, part, domain, subunit or configuration. The invention also provides an invention comprising a single variable domain (ISVD) that binds to a matrix metalloproteinase (MMP) and another single variable domain (ISVD) that binds to a cartilage proteoglycan, preferably aggrecan. of peptides. Another aspect concerns the polypeptide according to the invention for use as a medicament. Yet another aspect relates to a method of treating or preventing a disease or disorder in an individual, for example involving MMP13 activity, the method comprising administering to the individual an amount of a polypeptide according to the invention effective to treat or prevent symptoms of the disease or disorder. Other aspects, advantages, applications and uses of polypeptides and compositions will become clear from further disclosure herein. Throughout this specification a number of documents are referenced. Every document cited herein, whether above or below (including all patents, patent applications, scientific publications, manufacturer's instructions, teachings, etc.), is hereby incorporated by reference in its entirety. Nothing contained herein should be construed as an admission that the present invention is not entitled to antedate the date of these disclosures by virtue of prior invention.

There is still a need for safe and effective OA agents. Such agents should comply with various and often competing requirements, especially when envisioned in a broadly applicable format. Rather, the format should ideally be usable by a wide range of patients. The format should preferably be safe and not induce infection due to frequent IA administration. In addition, this format is more patient-friendly. For example, the format should have an extended half-life in the joint such that the format is not removed immediately upon administration. However, extending half-life should preferably not introduce off-target activity and side effects or limit efficacy. The present invention realizes at least one of these requirements. Based on unconventional screening, characterization and combination strategies, the inventors were surprised to observe that immunoglobulin single variable domains (ISVD) performed exceptionally well in vitro and in vivo experiments. Furthermore, the inventors were able to reconstruct ISVD to outperform the comparator drugs. In dual-parallel mode, this performance is not only preserved but even improved. On the other hand, it is also shown that the ISVD of the present invention is significantly more effective than the comparative molecules. The present invention provides polypeptides that antagonize MMPs, particularly MMP13, with improved prophylactic, therapeutic and/or pharmacological properties compared to comparative molecules, including a safer profile. Therefore, the present invention relates to ISVDs and polypeptides directed against MMPs and/or that can specifically bind to MMPs (as defined herein) and modulate their activity. Preferably, the MMPs are selected from the group consisting of: MMP13 (collagenase) ), MMP8 (collagenase), MMP1 (collagenase), MMP19 (matrix metalloproteinase RASI) and MMP20 (ameliolysin). Preferably, the MMP is MMP13. In particular, the polypeptide includes at least one immune protein that specifically binds to MMP13. Globulin single variable domain (ISVD), in which binding to MMP13 modulates the activity of MMP13. Definitions Unless otherwise indicated or defined, all terms used have their customary meanings in the art and will be understood by those skilled in the art. Reference is made to, for example, standard manuals such as Sambrook et al. (Molecular Cloning: A Laboratory Manual (2^nd Ed.) Vols. 1-3, Cold Spring Harbor Laboratory Press, 1989), F. Ausubel et al. (Current protocols in molecular biology, Green Publishing and Wiley Interscience, New York, 1987), Lewin (Genes II, John Wiley & Sons, New York, N.Y., 1985), Old et al. (Principles of Gene Manipulation: An Introduction to Genetic Engineering (2nd edition) University of California Press, Berkeley, CA, 1981), Roitt et al. (Immunology (6^th Ed.) Mosby/Elsevier, Edinburgh, 2001), Roitt et al (Roitt’s Essential Immunology (10^th Ed.) Blackwell Publishing, UK, 2001) and Janeway et al. (Immunobiology (6^th Ed.) Garland Science Publishing/Churchill Livingstone, New York, 2005), and the general background cited in this article. Unless otherwise instructed, all methods, steps, techniques and operations not specifically described in detail can be performed and have been performed in a manner known per se, as would be apparent to a person skilled in the art. Reference is further made to, for example, standard manuals and the general background mentioned herein and other references cited herein; and to, for example, the comments below: Presta (Adv. Drug Deliv. Rev. 58(5-6): 640-56, 2006 ), Levin and Weiss (Mol. Biosyst. 2(1): 49-57, 2006), Irving et al. (J. Immunol. Methods 248(1-2): 31-45, 2001), Schmitz et al. (Placenta 21 Suppl. A: S106-12, 2000), Gonzales et al. (Tumour Biol. 26(1): 31-43, 2005), which describe techniques for protein engineering (such as affinity maturation) and improved proteins (such as immunoglobulin Other techniques for specificity and other desired properties of proteins. It should be noted that as used herein, the singular forms "a", "an" and "the" include plural references unless the context clearly indicates otherwise. Thus, for example, reference to "a reagent" includes one or more of these different reagents and reference to "the method" includes those known to those of ordinary skill in the art to be Equivalent steps and methods described that are modified or substituted for use in the methods described herein. Unless otherwise indicated, it is understood that the term "at least" preceding a series of elements refers to each element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is intended that this invention encompasses such equivalents. The term "and/or" used at any time herein includes the meaning of "and", "or" and "all or any other combination of elements connected by that term". As used herein, the term "about" or "approximately" means within 20%, preferably within 15%, and more preferably within 10% of a given value or range. , and the best is within 5%. Throughout this specification and the patent claims that follow, it is understood that the word "comprise" and variations (such as "comprises" and "comprising") mean, unless the context requires otherwise. Includes a stated integer or step, or group of integers or steps, but does not exclude any other integer, step, or group of integers or steps. When used herein, the term "comprising" may be replaced by the term "containing" or "including", or sometimes, when used herein, by the term "having". The term "sequence" as used herein should generally be understood (e.g. in terms such as "immunoglobulin sequence", "antibody sequence", "variable domain sequence", "V_HH The term "sequence" or "protein sequence") includes both the related amino acid sequences and the nucleic acid or nucleotide sequences encoding them, unless the context requires a more limited interpretation. Amino acid sequences are construed to mean A single amino acid or an unbranched sequence of two or more amino acids, depending on the context. Nucleotide sequence is interpreted to mean an unbranched sequence of 3 or more nucleotides. Amino group The acids are those L-amino acids commonly found in naturally occurring proteins. Amino acid residues are represented according to the standard three-letter or one-letter amino acid codes. See, for example, Table A- on page 48 of WO 08/020079 2. Those amino acid sequences containing D-amino acids are not intended to be encompassed by this definition. Any amino acid sequence containing a post-translationally modified amino acid may be described as having the modifications shown in this Table A-2 The original translation (e.g., hydroxylation or glycation) of the amino acid sequence of the position symbol, but such modifications should not be explicitly shown in the amino acid sequence. This definition covers linkages, cross-links, and end caps that can be sequence modified , any peptide or protein represented by non-peptide bonds, etc. The terms "protein", "peptide", "protein/peptide" and "polypeptide" are used interchangeably throughout this disclosure and for the purposes of the present invention. Have the same meaning. Each term refers to an organic compound composed of a straight chain of two or more amino acids. The compound may have 10 or more amino acids, 25 or more amino acids, 50 or More amino acids, 100 or more amino acids, 200 or more amino acids, and even 300 or more amino acids. Those skilled in the art understand that polypeptides generally contain less than proteins Amino acids, although there is no technically recognized cut-off point for the number of amino acids that distinguishes polypeptides from proteins; polypeptides can be produced by chemical synthesis or recombinant methods; and proteins are usually produced by recombinant methods in vitro or in vivo, as in this It is known in the art that the amide bonds in the primary structure of polypeptides are conventionally written in the order of amino acids, with the amine terminus (N-terminus) of the polypeptide always on the left and the acid terminus (C-terminus) always on the right. For example, when the nucleic acid or amino acid sequence has been associated with at least one other component ( When separated from another nucleic acid, another protein/polypeptide, another biological component or macromolecule, or at least one contaminant, impurity or minor component), a nucleic acid or amino acid sequence is considered to be ) is isolated (form)". In particular, a nucleic acid or amino acid sequence is considered to have been purified at least 2-fold, in particular at least 10-fold, more in particular at least 100-fold and up to 1000-fold or more. The amino acid sequence is "(substantially) isolated." The nucleic acid or amino acid "in (substantially) isolated form" is preferably substantially homogeneous, as determined using a suitable technique, such as suitable chromatography techniques, such as polyacrylamide gel electrophoresis. When a nucleotide sequence or an amino acid sequence, respectively, is said to "comprise" another nucleotide sequence or amino acid sequence or "consist essentially of" another nucleotide sequence or amino acid sequence, this may mean that the later-mentioned nucleotide sequence or amino acid sequence has been incorporated into the first-mentioned nucleotide sequence or amino acid sequence, respectively, but more Frequently, this usually means that the first-mentioned nucleotide sequence or amino acid sequence respectively contains a stretch of nucleotides or amino acid residues within its sequence, and has the same nucleotide sequence as the later-mentioned sequence respectively. or amino acid sequence, regardless of how the aforementioned sequence is actually generated or obtained (which may, for example, be by any suitable method described herein). By way of non-limiting example, when a polypeptide of the invention is said to comprise an immunoglobulin single variable domain ("ISVD"), this may mean that the immunoglobulin single variable domain sequence has been incorporated into the polypeptide of the invention in the sequence, but more often, this generally means that the polypeptide of the invention contains within its sequence the sequence of an immunoglobulin single variable domain, regardless of how the polypeptide of the invention is produced or obtained. Likewise, when a nucleic acid or nucleotide sequence is said to comprise another nucleotide sequence, the first-mentioned nucleic acid or nucleotide sequence is preferably such that when expressed as an expression product (e.g., a polypeptide), the later-mentioned nucleic acid or nucleotide sequence is and the amino acid sequence encoded by the nucleotide sequence forms part of the expression product (in other words, the nucleotide sequence mentioned later is in the same reading frame as the larger nucleic acid or nucleotide sequence mentioned earlier ). Likewise, when a construct of the invention is said to comprise a polypeptide or ISVD, this may mean that the construct comprises at least the polypeptide or ISVD respectively, but more often it means that the construct comprises in addition to the polypeptide or ISVD. In addition, there are groups, residues (such as amino acid residues), moieties and/or binding units, regardless of how the groups, residues (such as amino acid residues), moieties and / or combined units, regardless of how the construct was created or obtained. By "consisting essentially of" it is meant that the immunoglobulin single variable domain used in the method of the invention is completely identical to or corresponds to the immunoglobulin single variable domain of the invention. Domains have a limited number of amino acid residues added on the amine terminus, the carboxyl terminus, or both the amine terminus and the carboxyl terminus (such as 1 to 20 amino acid residues, e.g., 1 to 10 amino acid residues) residues, and preferably 1 to 6 amino acid residues, such as 1, 2, 3, 4, 5 or 6 amino acid residues) of the immunoglobulin single variable domain of the invention. For the purpose of comparing two or more nucleotide sequences, the percent "sequence identity" between a first nucleotide sequence and a second nucleotide sequence can be determined by [in the first nucleotide sequence The number of nucleotides identical to the nucleotide at the corresponding position in the second nucleotide sequence] divided by [the total number of nucleotides in the first nucleotide sequence] and multiplied by [100%] Calculation in which each deletion, insertion, substitution or addition of a nucleotide in a second nucleotide sequence is considered a difference in a single nucleotide (position) compared to the first nucleotide sequence. Alternatively, the degree of sequence identity between two or more nucleotide sequences can be calculated using known computer algorithms for sequence alignment (such as NCBI Blast v2.0) and using standard settings. . Some other techniques, computer algorithms and settings for determining the degree of sequence identity are described in, for example, WO 04/037999, EP 0967284, EP 1085089, WO 00/55318, WO 00/78972, WO 98/49185 and GB 2357768. For purposes of determining the percent "sequence identity" between two nucleotide sequences according to the calculation method outlined above, the nucleotide sequence with the highest number of nucleotides is often considered the "first" nucleotide. sequence and other nucleotide sequences are considered the "second" nucleotide sequence. For the purpose of comparing two or more amino acid sequences, "sequence identity" between a first amino acid sequence and a second amino acid sequence (also referred to herein as "amino acid identity" ) percentage can be calculated by dividing [the number of amino acid residues in the first amino acid sequence that are identical to the amino acid residue at the corresponding position in the second amino acid sequence] by [the number of amino acid residues in the first amino acid sequence] total number of amino acid residues in the acid sequence] and multiplied by [100%] to calculate where each amino acid residue in the second amino acid sequence is missing compared to the first amino acid sequence , insertion, substitution or addition is considered to be a difference in a single amino acid residue (position), that is, an "amino acid difference" as defined herein. Alternatively, the degree of sequence identity between two amino acid sequences can be determined using known computer algorithms for determining the degree of sequence identity of nucleotide sequences (such as those mentioned above), Calculations are performed as usual using standard settings. For purposes of determining the percent "sequence identity" between two amino acid sequences according to the calculation method outlined above, the amino acid sequence with the highest number of amino acid residues is often considered the "first" amine The amino acid sequence and other amino acid sequences are regarded as the "second" amino acid sequence. Furthermore, when determining the degree of sequence identity between two amino acid sequences, one skilled in the art may consider so-called "conservative" amino acid substitutions, which are generally described as in which the amino acid residue has a chemical structure An amino acid substitution that is similar to the substitution of another amino acid residue and has little or essentially no effect on the function, activity, or other biological properties of the polypeptide. Such conservative amino acid substitutions are well known in the art, such as WO 04/037999, GB 335768, WO 98/49185, WO 00/46383 and WO 01/09300; and these substitutions are (preferred) Types and/or combinations may be selected based on relevant teachings from, for example, WO 04/037999 and WO 98/49185 and from further references cited herein. Preferably these conservative substitutions are substitutions in which an amino acid within each of the following groups (a) to (e) is replaced by another amino acid residue within the same group: (a) small aliphatic, non- Polar or slightly polar residues: Ala, Ser, Thr, Pro and Gly; (b) Polar, negatively charged residues and their (uncharged) amides: Asp, Asn, Glu and Gln; (c) Polar, negatively charged residues Positively charged residues: His, Arg and Lys; (d) large aliphatic, non-polar residues: Met, Leu, Ile, Val and Cys; and (e) aromatic residues: Phe, Tyr and Trp. Particularly preferred conservative substitution systems are as follows: Ala becomes Gly or becomes Ser; Arg becomes Lys; Asn becomes Gln or becomes His; Asp becomes Glu; Cys becomes Ser; Gln becomes Asn; Glu becomes Asp; Gly becomes Ala or becomes Pro; His becomes Asn or becomes Gln; Ile becomes Leu or becomes Val; Leu becomes Ile or becomes Val; Lys becomes Arg, becomes Gln or becomes Glu; Met becomes Leu, becomes Tyr or becomes Ile; Phe becomes Met, becomes Leu or becomes Tyr ; Ser becomes Thr; Thr becomes Ser; Trp becomes Tyr; Tyr becomes Trp; and/or Phe becomes Val, becomes Ile or becomes Leu. Any amino acid substitutions applied to the polypeptides described herein may also be based on amino acid variations between homologous proteins of different species developed by Schulz et al. ("Principles of Protein Structure", Springer-Verlag, 1978) Analysis of frequencies, analysis of structure-forming potential developed by Chou and Fasman (Biochemistry 13: 211, 1974; Adv. Enzymol., 47: 45-149, 1978), and analysis of structure-forming potential by Eisenberg et al. (Proc. Natl. Acad Sci. USA 81: 140-144, 1984), Kyte and Doolittle (J. Molec. Biol. 157: 105-132, 1981) and Goldman et al. (Ann. Rev. Biophys. Chem. 15: 321-353, 1986 ), the entire text of which is incorporated herein by reference. Information on the primary, secondary and tertiary structures of Nanobodies is given in the Summary of the Invention herein and in the general background cited above. Also for this purpose, V from Llama_HH The crystal structure of the domain is provided by, for example, Desmyter et al. (Nature Structural Biology, 3: 803, 1996), Spinelli et al. (Natural Structural Biology, 3: 752-757, 1996), and Decanniere et al. (Structure, 7(4): 361, 1999) given. About V in Xi Zhi_H V is formed in the domain_H /V_L Additional information on some of the amino acid residues at the interface and possible camelizing substitutions at these positions can be found in the prior art cited above. An amino acid sequence and a nucleic acid sequence are said to be "identical" if they have 100% sequence identity (as defined herein) over their entire length. When comparing two amino acid sequences, the term "amino acid difference" refers to the insertion, deletion or substitution of a single amino acid residue at a position in the first sequence compared to the second sequence; it should be understood that both The amino acid sequence may contain one, two or more such amino acid differences. More specifically, in the ISVD and/or polypeptide of the present invention, the term "amino acid difference" refers to the difference in b), d) or f) compared to the CDR sequence of a), c) or e) respectively. Insertion, deletion or substitution of a single amino acid residue at a specified CDR sequence position; it is understood that the CDR sequences of b), d) and f) are comparable to the CDR sequences of a), c) or e) respectively. Containing one, two, three, four or up to five of these amino acid differences. An "amino acid difference" may be any one, two, three, four or up to five substitutions, deletions or insertions or any combination thereof, which improves or at least improves the properties of the MMP13 binding agents of the invention (such as the polypeptides of the invention). Unduly impairs a desired property or balance or combination of desirable properties of a MMP13 binding agent of the invention, such as a polypeptide of the invention. In this regard, the resulting MMP13 binding agents of the invention (such as the polypeptides of the invention) should be at least Binds to MMP13 with the same, approximately the same or higher affinity. Affinity can be measured by any suitable method known in the art, but is preferably measured as described in the Examples paragraph. In this regard, the amino acid sequences of the CDRs according to b), d) and/or f) may be individually self-isolated by means of affinity maturation using one or more affinity maturation techniques known per se or as described in the examples. Amino acid sequences derived from the amino acid sequences of a), c) and/or e). For example, and depending on the host organism in which the polypeptides of the invention are expressed, such deletions and/or substitutions may remove one or more sites for post-translational modification (such as one or more glycation sites) This method of design is within the capabilities of those skilled in the art (refer to the embodiment). In the context of any SEQ ID NO, "represented by" as used herein is equivalent to "comprising or consisting of "the SEQ ID NO", and is preferably equivalent to "the SEQ ID NO". ID NO〞Composed of〞. As used in this manual, "Nanobody family", "V_HH "Family" or "Family" refers to Nanobodies and/or V that have the same length._HH Sequence groups (i.e., having the same number of amino acids within their sequences) and their amino acid sequences between positions 8 and 106 (according to Kabat numbering) have 89% or higher amino acids Sequence identity. The terms "antigenic region" and "antigenic determinant" are used interchangeably to refer to an antigen-binding molecule (such as an immunoglobulin of the invention, a conventional antibody, an immunoglobulin single variable domain and/or a polypeptide), and More particularly a portion of a giant molecule (such as a polypeptide or protein) recognized by the antigen binding site of the molecule. The epitope defines the minimum binding site for an immunoglobulin and therefore represents the target for immunoglobulin specificity. The part of an antigen-binding molecule (such as the immunoglobulin of the present invention, a conventional antibody, an immunoglobulin single variable domain and/or a polypeptide) that recognizes an antigenic determinant is called a "paratope." Can "bind" or "specifically bind", "have affinity to" and/or "have specificity" to a specific epitope, antigen or protein (or at least a part, fragment or epitope thereof) An amino acid sequence (such as an immunoglobulin single variable domain of the invention, an antibody, a polypeptide or generally an antigen or fragment thereof that binds to a protein or polypeptide) is said to be "against" or "directed against" )"the epitope, antigen or protein, or is a "binding" molecule related to such epitope, antigen or protein, or is said to be an "anti" epitope, "anti" antigen or "anti" protein ( For example "anti" MMP13). Affinity represents the strength or stability of molecular interactions. Affinity is often expressed in units of moles/liter (or M)._D or the dissociation constant is given. Affinity can also be associated with the association constant K_A means that it is equal to 1/K_D and has (mol/liter)^-1 (or M^-1 ) unit. In this specification, the stability of the interaction between two molecules is mainly determined by the K of their interaction._D Expressed in the form of value; those familiar with this technology will understand that in view of K_A =1/K_D relationship, with its K_D The strength of the molecular interaction specified by the value can also be used to calculate the corresponding K_A value. K_D The value is also characterized in a thermodynamic sense by the strength of molecular interactions, since it is determined by the well-known relationship DG=RTln(K_D )(equivalent to DG= -RTln(K_A )) is related to the change in free energy of binding (DG), where R is equal to the gas constant, T is equal to the absolute temperature and ln represents the natural logarithm. The K of biological interactions that are considered meaningful (e.g., specific)_D Usually tied at 10^-12 M(0.001 nM) to 10^-5 M (10000 nM). The stronger the interaction, the K_D The lower. K_D The dissociation rate constant of the complex (in terms of k_off expressed) and its association rate (expressed as k_on expressed) expressed by the ratio of expressed (such that K_D =k_off /k_on and K_A =k_on /k_off ). Dissociation rate k_off has unit s^-1 (where s is the SI unit notation of seconds). Association rate k_on Has unit M^-1 s^-1 . The association rate can be between 10² M^-1 s^-1 to about 10⁷ M^-1 s^-1 changes between them, approaching the diffusion-limited association rate constant of bimolecular interactions. The dissociation rate is related to t_1/2 =ln(2)/k_off The half-lives of the given molecular interactions are related. The dissociation rate can be between 10^-6 s^-1 (in terms of multiple days t_1/2 Approximately irreversible complex) to 1 s^-1 (t_1/2 =0.69 s). Specific binding of an antigen-binding protein (such as ISVD) to an antigen or antigenic determinant may be determined in any suitable manner known per se, including, for example, saturation binding assays and/or competitive binding assays, such as radioimmunoassays (RIA) ), enzyme immunoassays (EIA) and sandwich competition assays, and their different variations known per se in this technology; and other technologies mentioned herein. The affinity of a molecular interaction between two molecules can be measured via different techniques known per se, such as the well-known surface plasmon resonance (SPR) biosensor technology (see e.g. Ober et al. 2001, Intern. Immunology 13 : 1551-1559), in which a molecule is fixed on the biosensor chip and other molecules pass through the fixed molecule under flow conditions to obtain k_on , k_off Measure and therefore get K_D (or K_A )value. This can be performed, for example, using the well-known BIACORE® instrument (Pharmacia Biosensor AB, Uppsala, Sweden). The kinetic exclusion assay (KINEXA®) (Drake et al. 2004, Analytical Biochemistry 328: 35-43) measures binding events in solution without labeling the binding partner and is based on kinetic exclusion complex dissociation. In-solution affinity analysis can also be performed using the GYROLAB® Immunoassay System, which provides a platform for automated bioanalysis and rapid sample transfer (Fraley et al. 2013, Bioanalysis 5:1765-74). Those skilled in the art will also understand that if the measurement method affects the inherent binding affinity of the underlying molecule in some way, such as distortion associated with a coating on a biosensor of a molecule, then the measured K_D May be equivalent to visual K_D . Likewise, if a molecule contains more than one recognition site for other molecules, it is possible to measure the visual K_D . In such cases, the measured affinity can be affected by the affinity of the interaction between the two molecules. In particular, accurate measurement of K_D It can be very laborious, and therefore the visual K_D value to evaluate the binding strength of two molecules. It should be noted that visual K can be used as long as all measurements are performed in a consistent manner (e.g. keeping calibration conditions constant)._D measure as true K_D is an approximate value, and therefore in this document K should be_D And as K_D viewed with equal importance or relevance. The term "specificity" refers to the number of different types of antigens or antigenic determinants that can bind to a particular antigen-binding molecule or antigen-binding protein molecule (such as an ISVD or polypeptide of the invention), for example in paragraph 53 of WO 08/020079 Go to paragraph n) on page 56. The specificity of an antigen-binding protein can be based on affinity and/or retention assays, as described on pages 53 to 56 of WO 08/020079 (incorporated herein by reference), which also describes some methods for measuring antigen binding. Preferred techniques for binding between molecules (such as polypeptides of the invention or ISVD) and coherent antigens. Antigen-binding proteins (such as ISVDs and/or polypeptides of the invention) are usually expressed as 10^-5 to 10^-12 Mol/liter or less, and preferably 10^-7 to 10^‑12 Mol/liter or less, and preferably 10^-8 to 10^-12 Dissociation constant in moles/liter (K_D )(i.e. 10⁵ to 10¹² Liter/mol or higher, preferably 10⁷ to 10¹² Liter/mol or higher, and preferably 10⁸ to 10¹² Liter/mol association constant (K_A )) binds to those antigens. It is generally considered to be greater than 10^-4 Mol/L of any K_D value (or less than 10⁴ Liter/mol of any K_A value) indicates nonspecific binding. Preferably the monovalent ISVD of the invention has an affinity of less than 500 nM, preferably less than 200 nM, more preferably less than 10 nM, such as less than 500 pM, such as between 10 and 5 pM or less. Binds to desired antigen. When the immunoglobulin single variable domain and/or polypeptide binds to the first antigen with affinity (as described above, and appropriately expressed as K_D Value, K_A Value, K_off Velocity and/or K_on rate) is at least 10 times, such as at least 100 times, and preferably at least 1000 times or more, better than the affinity of the immunoglobulin single variable domain and/or polypeptide for binding to the second target or antigen. A single variable domain of a protein and/or a polypeptide is said to be "specific" for a (first) target or antigen relative to another (second) target or antigen. For example, the K of an immunoglobulin single variable domain and/or polypeptide that binds to the first target or antigen_D The value is greater than the K of the immunoglobulin single variable domain and/or polypeptide binding to the second target or antigen._D At least 10 times smaller, such as at least 100 times smaller, and preferably at least 1000 times smaller or even smaller. Preferably when an immunoglobulin single variable domain and/or polypeptide is "specific" for a first target or antigen versus a second target or antigen, it is directed against (as defined herein) the first target or antigen. target or antigen, but is not directed against the second target or antigen. Specific binding of an antigen-binding protein to an antigen or antigenic determinant may be determined in any suitable manner known per se, including, for example, saturation binding assays, Scatchard analysis, and/or competitive binding assays, such as radioimmunoassay. Assays (RIA), enzyme immunoassays (EIA), and sandwich competitive assays, and their different variations known in this technology; and other technologies mentioned herein. As will be appreciated by those skilled in the art and as described on pages 53 to 56 of WO 08/020079, the dissociation constant may be a real or an apparent dissociation constant. Methods for determining dissociation constants are known to those skilled in the art and include, for example, the techniques mentioned on pages 53 to 56 of WO 08/020079. Another preferred method that can be used to assess affinity is the 2-step ELISA (enzyme-linked immunosorbent assay) procedure of Friguet et al. in 1985 (J. Immunol. Methods 77: 305-19). This method establishes solution-phase binding equilibrium measurements and avoids possible distortions associated with adsorption of one of the molecules on the support (such as plastic). The dissociation constant may be a real or an apparent dissociation constant, as will be apparent to those skilled in the art and as described, for example, on pages 53 to 56 of WO 08/020079. Methods for determining dissociation constants are known to those skilled in the art and include, for example, the techniques mentioned on pages 53 to 56 of WO 08/020079. In one aspect, the invention relates to MMP13 binding agents, such as the ISVDs and polypeptides of the invention, wherein the MMP13 binding agent does not bind to MMP1 or MMP14 (membrane type). Finally, it should be noted that the experienced scientist may in many cases determine that it is convenient to determine the binding affinity relative to some reference molecule. For example, to assess the strength of the binding between molecules A and B, one can e.g. Choose) easily detectable biotin) or other forms (fluorophore for fluorescence detection, chromophore for light absorption detection, biotin for streptavidin-mediated ELISA detection) ) appropriately labeled reference molecule C. Usually the reference molecule C is kept at a fixed concentration and the A concentration is varied for a given concentration or amount of B. As a result, the IC corresponding to the concentration of A is obtained₅₀ value, which is halved by the signal measured with C in the absence of A. If K is known_{D ref(} Reference molecule K_D ) and the total concentration c of the reference molecule_ref , then it can be obtained from the following formula: K_D =IC₅₀ /(1+c_ref /K_Dref ) to obtain the view K of the interaction A-B_D . It should be noted that if c_ref ＜＜K_{D ref} , then K_D »IC₅₀ . If in a consistent manner (e.g. keeping a fixed c_ref ) to perform IC of binding agents for comparison₅₀ measurement, then differences in the strength or stability of molecular interactions can be measured by comparing IC₅₀ to evaluate and determine that this measure is equivalent to K throughout the article_D Or depending on K_D . Half of the maximum inhibitory concentration (IC₅₀ ) may also be a measure of the effectiveness of a compound in inhibiting biological or biochemical functions (eg, pharmacological effects). This quantitative measure represents how much polypeptide or ISVD (e.g., Nanobody) is required to inhibit half of a given biological process (or component of a process, i.e., enzyme, cell, cell receptor, chemotaxis, regression, metastasis, invasion, etc.). In other words, it is half (50%) of the maximum inhibitory concentration (IC) of the substance (50% IC or IC₅₀ ). IC of antagonists provided by the present invention (such as polypeptides or ISVD (such as nanobodies))₅₀ The value can be calculated by determining the concentration required to inhibit half of the maximum biological response of the agonist. Drug K_D This can be determined by constructing a dose-response curve and examining the effect of different concentrations of an antagonist, such as a polypeptide of the invention or an ISVD (eg, a Nanobody), on reversing agonist activity. Term half the maximum effective concentration (EC₅₀ ) means the concentration of a compound that induces a half-way response between baseline and maximum after a specified exposure time. In the context of the present invention, the EC₅₀ Used as a measure of the potency of a polypeptide or ISVD (eg Nanobody). EC for graded dose-response curves₅₀ Represents the concentration of a compound at which 50% of its maximum effect is observed. Concentrations are preferably expressed in molar units. In biological systems, small changes in ligand concentration often cause rapid response changes that follow a sigmoid function. The inflection point at which the reaction begins to slow down with increasing ligand concentration is the EC₅₀ . This can be determined mathematically by deriving a line of best fit. In most cases, it is convenient to rely on graphs for estimation. If EC is provided in the Examples paragraph₅₀ , then design experiments that reflect KD as accurately as possible. In other words, E.C.₅₀ The value can be considered as the KD value. The term "average KD" refers to the average KD value obtained in at least 1, but preferably more than 1, such as at least 2 experiments. The term "average" refers to the mathematical term "average" (the sum of data divided by the number of items in the data). It’s also about IC₅₀ , which is a measure of a compound's inhibitory effect (50% inhibition). IC₅₀ It is the most common summary measure for dose-response curves in competitive binding assays and functional antagonist assays. The most common summary measure for agonist/stimulant assays is EC₅₀ . The inhibition constant Ki is an indicator of how effective an inhibitor is; it is the concentration required to produce half of maximum inhibition. with IC₅₀ Different, IC₅₀ As may vary depending on the experimental conditions (but see above), Ki is an absolute value and is often referred to as the inhibition constant of the drug. Suppression constant K_i It can be calculated using the Cheng-Prusoff formula:where [L] is the fixed ligand concentration. As the term is used herein, "potency" of a polypeptide of the invention is a function of the amount of the polypeptide of the invention required to produce its specific effect. It is based on the IC of the polypeptide₅₀ The reciprocal of is simply measured. Potency refers to the ability of the polypeptide of the invention to modulate and/or partially or completely inhibit the activity of MMP13. More specifically, potency may refer to the ability of the polypeptide to reduce or even completely inhibit MMP13 activity as defined herein. Specifically, potency may refer to the polypeptide's ability to inhibit proteolysis (such as protease activity, endopeptidase activity) and/or substrates (such as aggrecan, collagen II, collagen I, collagen III, collagen IV, collagen protein IX, collagen X, collagen XIV and gelatin) binding ability. Potency can be measured by any suitable assay known in the art or described herein. The "efficacy" of the polypeptide of the present invention is measured at the maximum intensity of the effect itself at a saturated polypeptide concentration. Efficacy refers to the maximum response that can be achieved from a polypeptide of the invention. Efficacy refers to the ability of a polypeptide to achieve the desired (therapeutic) effect. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide is expressed between 1E^-07 M and 1E^-13 K between M_D Binds to MMP13, such as between 1E^-08 M and 1E^-12 Between M, preferably at most 1E^-07 M, preferably less than 1E^-08 M or 1E^-09 M, or even below 1E^‑10 M, such as 5E^-11 M, 4E⁼¹¹ M, 3E^‑11 M, 2E^-11 M, 1.7E^-11 M, 1E^‑11 M, or even 5E^-12 M, 4E^-12 M, 3E^‑12 M, 1E^-12 M, which is measured, for example, with KinExA. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide is expressed between 1E^-07 M and 1E^-12 IC between M₅₀ Inhibit MMP13 activity, such as between 1E^-08 M and 1E^-11 M, which is determined, for example, by a competitive ELISA, a competitive TIMP-2 ELISA, a fluorescent peptide assay, a fluorescent collagen assay or a collagen lysis assay, such as detailed in the Examples paragraph. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide is expressed with at most 1E^-07 M of IC₅₀ Inhibit the activity of MMP13, preferably 1E^-08 M, 5E^-09 M or 4E^-9 M, 3E^-9 M, 2E^-9 M, such as 1E^-9 M. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide is expressed between 1E^-07 M and 1E^-12 EC between M₅₀ Binds to MMP13, such as between 1E^-08 M and 1E^-11 M, for example, it is measured by ELISA, competitive TIMP-2 ELISA, fluorescent peptide assay, fluorescent collagen assay or collagen dissolution assay. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide is expressed at less than 5E^-04 (s^-1 ) binds to MMP13, which is measured, for example, by SPR. An amino acid sequence (such as an ISVD or polypeptide) is said to be specific for two different antigens or antigenic determinants (such as from different species) if it is specific (as defined herein) for these different antigens or antigenic determinants. MMP13s of mammals, such as human MMP13, dog MMP13, bovine MMP13, rat MMP13, porcine MMP13, mouse MMP13, rabbit MMP13, cynomolgus monkey MMP13 and/or rhesus monkey MMP13) have "cross-reactivity". It is understood that ISVDs or polypeptides may be considered to be cross-reactive, although the binding affinities for two different antigens may be different, such as by a factor of 2, 5, 10, 50, 100, or even greater, provided that the binding affinities for these different antigens are The antigen or antigenic determinant is specific (as defined herein). MMP13 is also known as CLG3 or collagenase 3, MANDP1, MMP-13, matrix metallopeptidase 13 or MDST. Information about the related structure of MMP13 can be found, for example, by the UniProt accession number described in Table 1 below (see Table B)."Human MMP13" refers to MMP13 comprising the amino acid sequence of SEQ ID NO:115. In one aspect, the polypeptide of the present invention specifically binds to MMP13 from Homo sapiens, mouse, wolf, domestic cow, rhesus macaque, brown rat, chicken and/or chimpanzee, preferably from human MMP13, more preferably from human MMP13. Preferably SEQ ID NO:115. Terms "(cross)-block ((cross)-block, (cross)-blocked, (cross)-blocking)", "competitive binding", "(cross) competition ((cross)-compete, (cross)- Competing and (cross)-competition) are used interchangeably herein to mean that an immunoglobulin, antibody, ISVD, polypeptide or other binding agent interferes with other immunoglobulins, antibodies, ISVDs, polypeptides or binding agents with a given target. Target binding ability. The extent to which an immunoglobulin, antibody, ISVD, peptide or other binding agent is able to interfere with the binding of another to the target, and therefore whether it can be said to be cross-blocking in accordance with the present invention, can be determined using competitive binding assays commonly known in the art. The purified ISVD can be assayed, such as, for example, by screening purified ISVD against ISVD expressed on phage in a competitive ELISA, as described in the Examples. For determining whether an immunoglobulin, antibody, immunoglobulin single variable domain, polypeptide or other binding agent (cross) blocks, is able to (cross) block, competitively binds or (cross) competes against a target ( (as defined herein) are described, for example, in Xiao-Chi Jia et al. (Journal of Immunological Methods 288: 91-98, 2004), Miller et al. (Journal of Immunological Methods 365: 118-125, 2011) and/or herein in the methods described (see, e.g., Example 7). The present invention relates to polypeptides as described herein, such as SEQ ID NOs: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20, The polypeptide represented by 21, 22, 2, 3, 4, 5, 6, 7 or 8, wherein the polypeptide competes with the polypeptide, which is determined, for example, by a competitive ELISA. The present invention relates to assays with polypeptides as described herein (such as SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, A method of competing with a competitor (such as a polypeptide) represented by any one of 20, 21, 22, 2, 3, 4, 5, 6, 7 or 8), wherein a polypeptide as described herein is competed with a competitor (such as a polypeptide ) competes or cross-blocks its binding to MMP13, such as human MMP13 (SEQ ID NO: 115), wherein the competitor's binding to MMP13 in the absence of the polypeptide of the invention is compared to the binding of the competitor to MMP13 in the presence of the polypeptide of the invention. The combination with MMP13 is reduced by at least 5%, such as 10%, 20%, 30%, 40%, 50% or even more, such as 80%, 90% or even 100% (i.e. almost cannot be detected). Competition and cross-blocking can be determined in any manner known in the art, such as competition ELISA. In one aspect, the invention relates to a polypeptide of the invention, wherein the polypeptide cross-blocks with SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16 , at least one of the polypeptides represented by 17, 18, 19, 20, 21, 22, 2, 3, 4, 5, 6, 7 or 8 binds to MMP13 and/or is SEQ ID NO: 111, 11, 112, At least one of the polypeptides represented by 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 2, 3, 4, 5, 6, 7 or 8 One cross-blocks binding to MMP13. The invention also relates to polypeptides as described herein (such as SEQ ID NOs: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20 , 21, 22, 2, 3, 4, 5, 6, 7 or 8) a competitor, wherein the competitor competes with the polypeptide as described herein or cross-blocks binding to MMP13, wherein compared to the present invention The binding of the polypeptide to MMP13 in the absence of the competitor, the binding of the polypeptide of the invention to MMP13 in the presence of the competitor is reduced by at least 5%, such as 10%, 20%, 30%, 40%, 50% or even More, such as 80% or even more, such as at least 90% or even 100% (ie barely detectable in a given assay). In one aspect, the invention relates to polypeptides of the invention (such as SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 2, 3, 4, 5, 6, 7 or 8) cross-blocking polypeptides binding to MMP13, and/or SEQ ID NO: 111, 11, 112, At least one of 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 2, 3, 4, 5, 6, 7 or 8 crosses Blocking binding to MMP13, preferably wherein the polypeptide comprises at least one VH, VL, dAb, immunoglobulin single variable domain (ISVD) that specifically binds to MMP13, wherein binding to MMP13 modulates the activity of MMP13. "MMP13 activity" and "activity of MMP13" (these terms are used interchangeably herein) include, but are not limited to, proteolytic activity, such as protease activity (also known as protease or peptidase activity) and, on the other hand, endopeptidase activity, and binding to substrates, such as through thrombin-like domains and peptidoglycan-binding domains. MMP13 activity includes binding to and/or proteolysis of substrates such as aggrecan, collagen II, collagen I, collagen III, collagen IV, collagen IX, collagen X, collagen XIV, and gelatin . Proteolysis as used herein refers to the breaking of proteins into smaller polypeptides or amino acids by hydrolysis of the peptide bonds that link amino acids together in the polypeptide chain. In the context of the present invention, "modulating or to modulate" generally means changing activity by MMP13, as measured using suitable in vitro, cellular or in vivo assays (such as those mentioned herein). In particular, "modulating or to modulate" may mean reducing or inhibiting MMP13 activity, or alternatively increasing MMP13 activity, using suitable in vitro, cellular or in vivo assays such as those mentioned herein. and) measured, modulated by at least 1%, preferably at least 5%, such as at least 10% or at least 25, compared to the MMP13 activity in the same assay under the same conditions but not in the presence of an ISVD or polypeptide of the invention. %, such as at least 50%, at least 60%, at least 70%, at least 80% or 90% or more. Therefore, the present invention relates to a polypeptide as described herein, wherein the polypeptide modulates MMP13 activity, preferably inhibits MMP13 activity. Accordingly, the present invention relates to a polypeptide as described herein, wherein the polypeptide inhibits the protease activity of MMP13, such as inhibiting proteolysis of a substrate, such as aggrecan, collagen II, collagen I, collagen III, collagen IV , collagen IX, collagen X, collagen XIV and/or gelatin. Accordingly, the present invention relates to a polypeptide as described herein, wherein the polypeptide blocks the interaction of MMP13 with a substrate such as aggrecan, collagen II, collagen I, collagen III, collagen IV, collagen IX, collagen X, collagen XIV and/or gelatin), wherein the collagen is preferably collagen II. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide blocks at least 20% of the binding of MMP13 to collagen and/or aggrecan, such as at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or even higher, for example, by a competitive ELISA-based assay (see Howes et al. 2014 J. Biol. Chem. 289:24091–24101) Determination. In one aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide antagonizes or inhibits the activity of MMP13, such as (i) protease activity, preferably cleavage of aggrecan and/or collagen, wherein the Collagen is preferably collagen II; (ii) collagen binds to a thrombin-like domain. Accordingly, the present invention relates to a polypeptide as described herein, wherein the polypeptide inhibits the protease activity of MMP13, preferably by at least 5%, such as 10%, 20%, 30%, 40%, 50% or even more, such as At least 60%, 70%, 80%, 90%, 95% or even higher, determined by any suitable method known in the art, such as a competitive assay or as described in the Examples paragraph. ISVD Unless otherwise indicated, the terms "immunoglobulin" and "immunoglobulin sequence" are both used to include full-length antibodies, their respective chain, and all parts, domains or fragments thereof (including, but not limited to, antigen-binding domains or fragments, respectively, such as V_HH Domain or V_H /V_L domain). As used herein, the term "domain" (of a polypeptide or protein) refers to a protein structure that has folds that retain its tertiary structure, regardless of the rest of the protein. Domains are often responsible for individual functional properties of a protein, and in many cases can be added, removed, or transferred to other proteins without losing the function of the rest of the protein and/or the domain. The term "immunoglobulin domain" as used herein refers to a globular region of an antibody chain (such as a chain of a conventional 4-chain antibody or a heavy chain antibody) or a polypeptide consisting essentially of such a globular region. Immunoglobulin domains are characterized by that they retain the immunoglobulin fold characteristic of antibody molecules, which consists of a bilayer sandwich of approximately 7 antiparallel β strands arranged into two β-pleated plates, optionally with conservative Sexual disulfide bonds are stable. The term "immunoglobulin variable domain" as used herein means an immunoglobulin domain consisting essentially of four "framework regions", referred to in the art and hereafter as "framework region 1" or "framework region 1" respectively. FR1", "Framework Region 2" or "FR2", "Framework Region 3" or "FR3" and "Framework Region 4" or "FR4"; these framework regions are based on three "Complementary Determining Regions" or "CDR" Interrupts are referred to as "Complementary Determining Region 1" or "CDR1", "Complementary Determining Region 2" or "CDR2" and "Complementary Determining Region 3" or "CDR3" in this technology and below respectively. Therefore, the general structure or sequence of an immunoglobulin variable domain can be represented as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. The immunoglobulin variable domain confers the antibody's specificity for the antigen by carrying the antigen-binding site. In a preferred embodiment of all aspects of the invention, the immunoglobulin single variable domain (ISVD) according to the invention preferably consists of or consists essentially of 4 framework regions in the general structure as outlined above (FR1 to FR4 respectively) and three complementary determining regions CDR1, CDR2 and CDR3. Preferred framework sequences are summarized, for example, in Table A-2 below and may be used in the ISVD of the present invention. Preferably the CDRs described in Table A-2 match the respective framework regions of the same ISVD construct. The terms "immunoglobulin single variable domain" (abbreviated herein as "ISVD" or "ISV") and "single variable domain" used interchangeably are defined in which an antigen-binding site is present on a single immunoglobulin domain and molecules formed from single immunoglobulin domains. This distinguishes immunoglobulin single variable domains from "conventional" immunoglobulins or fragments thereof, in which two immunoglobulin domains (in particular two variable domains) interact to form an antigen-binding site. Typically in conventional immunoglobulins, the heavy chain variable domain (V_H ) and the light chain variable domain (V_L ) interact to form the antigen-binding site. In the latter case, V_H and V_L The complementarity determining regions (CDRs) of the two contribute to the antigen binding site, that is, a total of 6 CDRs are involved in the formation of the antigen binding site. In view of the above definition, a conventional 4-chain antibody (such as an IgG, IgM, IgA, IgD or IgE molecule known in the art) or a Fab fragment, F(ab') derived from such a conventional 4-chain antibody Antigen-binding domains of 2 fragments, Fv fragments (such as disulfide-linked Fv or scFv fragments) or diabodies (all known in the art) are generally not considered to be immunoglobulin single variable domains because in In these cases, the binding to the respective epitope of the antigen is usually not with a (single) immunoglobulin domain, but with a pair (associated) of immunoglobulin domains (such as light and heavy chain variable domains). ) occurs, that is, the V of the immunoglobulin domain co-binds with the epitope of the respective antigen._H -V_L to happen. In contrast, ISVD is able to specifically bind to the epitope of the antigen without pairing with additional immunoglobulin variable domains. The binding site of ISVD is based on a single V_HH ,V_H or V_L domain formation. Therefore, the antigen-binding site of ISVD is formed by no more than three CDRs. Specifically, a single variable domain may be a light chain variable domain sequence (e.g., V_L sequence) or a suitable fragment thereof; or a heavy chain variable domain sequence (e.g. V_H Sequence or V_HH sequence) or a suitable fragment thereof; as long as it can form a single antigen-binding unit (i.e., a functional antigen-binding unit consisting essentially of a single variable domain, such that the single antigen-binding domain does not need to interact with another variable domain to form a functional antigen-binding unit). In one embodiment of the invention, ISVD is a heavy chain variable domain sequence (e.g., V_H sequence); more specifically, the ISVD may be a heavy chain variable domain sequence derived from a conventional quadribody or a heavy chain variable domain sequence derived from a heavy chain antibody. For example, an ISVD may be a (single) domain antibody (or a peptide suitable for use as a (single) domain antibody), a "dAb" or a dAb (or a peptide suitable for use as a dAb) or a Nanobody (as defined herein and including but not limited to VHH); other single variable domains or any suitable fragments of any of them. In particular, the ISVD may be a Nanobody® (as defined herein) or a suitable fragment thereof. [Note: Nanobody® and Nanobodies® are registered trademarks of Ablynx N.V.]. For a general description of Nanobodies, reference is made to the further description below, as well as to the prior art cited herein, such as that described in WO 08/020079 (page 16). Also known as VHH, V_H H domain, VHH antibody fragment and "V of VHH antibody"_HH Domain" was originally described as the antigen-binding immunoglobulin (variable) domain of a "heavy chain antibody" (i.e., an "antibody without a light chain"; Hamers-Casterman et al. 1993 Nature 363: 446-448). Select the term " V_HH domain" to distinguish these variable domains from the heavy chain variable domains (referred to herein as "V_H domain" or "VH domain") and the light chain variable domain (which is referred to herein as the "VH domain") present in conventional 4-chain antibodies._L Domain" or "VL domain"). For further description of VHH and nanobodies, refer to the review article by Muyldermans (Reviews in Molecular Biotechnology 74: 277-302, 2001) and the following patent applications, which are mentioned as general background technology : WO 94/04678, WO 95/04079 and WO 96/34103 of the Vrije Universiteit Brussel; WO 94/25591, WO 99/37681, WO 00/40968, WO 00/43507, WO 00/65057, WO 01 of Unilever /40310, WO 01/44301, EP 1134231 and WO 02/48193; WO 97/49805, WO 01/21817, WO 03/035694, WO 03/054016 and WO 03/055527 of the Vlaams Instituut voor Biotechnologie (VIB); WO 03/050531 to Algonomics N.V. and Ablynx N.V.; WO 01/90190 to the National Research Council of Canada; WO 03/025020 (=EP 1433793) to the Institute of Antibodies; and WO 04/041867, WO 04/041862 to Ablynx N.V. , WO 04/041865, WO 04/041863, WO 04/062551, WO 05/044858, WO 06/40153, WO 06/079372, WO 06/122786, WO 06/122787 and WO 06/122825 and updates of Ablynx N.V. Multiple published patent applications. Reference is also made to further prior art mentioned in these applications and in particular to the reference list mentioned on pages 41 to 43 of the international application WO 06/040153. The list and references are incorporated herein by reference. As described in these references, Nanobodies (particularly VHH sequences and partially humanized Nanobodies) may be specifically identified with one or more "Hallmarks" "Residues" are characterized by the presence of one or more in the framework sequence. Nanobodies (including humanization and/or camelization of Nanobodies, as well as other modifications, parts or fragments, derivatives or "Nanobodies" Further descriptions of "antibody fusions"), multivalent constructs (including some non-limiting examples of linker sequences) and different modifications that increase the half-life of Nanobodies and their formulations can be found, for example, in WO 08/101985 and WO 08/ 142164 in. For a further general description of Nanobodies, reference is made to the prior art cited herein, such as that described in WO 08/020079 (page 16). In particular, the framework sequences present in the MMP13 binding agents of the invention (such as the ISVDs and/or polypeptides of the invention) may contain one or more marker residues (for example in WO 08/020079 (Tables A-3 to A- 8)), making the MMP13 binding agent of the present invention a nanobody. Some preferred but non-limiting examples of (suitable combinations of) these framework sequences will become apparent from further disclosure herein (see, eg, Table A-2). Usually nanobodies (especially V_HH sequences and partially humanized Nanobodies) may be characterized in particular by the presence of one or more "marker residues" in one or more of the framework sequences (e.g. in WO 08/020079, page 61, line 24 to Further explanation in line 3 of page 98). More specifically, the present invention provides MMP13 binding agents comprising at least one immunoglobulin single variable domain, which is an amino acid sequence having the following (general) structure: FR1-CDR1-FR2-CDR2 -FR3-CDR3-FR4 Where FR1 to FR4 refer to framework regions 1 to 4 respectively, and CDR1 to CDR3 refer to complementarity determining regions 1 to 3 respectively, and the ISVD: i) is the same as SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 2, 3, 4, 5, 6, 7 or 8 amine groups At least one of the acid sequences has an amino acid identity of at least 80%, more preferably 90%, and even more preferably 95% (see Table A-1), where for the purpose of determining the degree of amino acid identity, Amino acid residues forming the CDR sequence are ignored. In this regard, reference is also made to Table A-2, which sets forth the frame 1 sequences (SEQ ID NOs: 67 to 79), frame 2 sequences of the immunoglobulin single variable domains of SEQ ID NOs: 1 to 22 and 109 to 112 (SEQ ID NO:80 to 87 and 108), frame 3 sequence (SEQ ID NO: 88 to 99 and 113 to 114) and frame 4 sequence (SEQ ID NO: 100 to 104); or ii) as shown in Table A-2 Combinations of framework sequences described in; and wherein: iii) Preferably in the amino acid residues at positions 11, 37, 44, 45, 47, 83, 84, 103, 104 and 108 according to Kabat numbering One or more are selected from marker residues, such as those mentioned in Tables A-3 to A-8 of WO 08/020079. MMP13 binding agents of the present invention (such as ISVDs and/or polypeptides of the present invention) may also contain those described in the following U.S. provisional applications all titled "Improved Immunoglobulin Variable Domains" which are also pending. Specific mutations/amino acid residues: US 61/994552 filed on May 16, 2014; US 61/014,015 filed on June 18, 2014; US 62/040,167 filed on August 21, 2014 ; and US 62/047,560 applied on September 8, 2014 (all assigned to Ablynx N.V.). In particular, MMP13 binding agents of the invention (such as ISVDs and/or polypeptides of the invention) may suitably contain (i) K or Q at position 112; or (ii) K or Q at position 110 combined with at position 110 The combination of V at position 11; or (iii) T at position 89; or (iv) L at position 89 and K or Q at position 110; or (v) V at position 11 and L at position 89; or any suitable combination of (i) to (v). As also described in these U.S. provisional applications that are also pending, when the MMP13 binding agent of the present invention (such as the ISVD and/or polypeptide of the present invention) contains the above (i) to (v) (or its appropriate Mutation of one of the combinations): - the amino acid residue at position 11 is preferably selected from L, V or K (and most preferably V); and/or - the amino acid residue at position 14 The residue is preferably selected from A or P; and/or - the amino acid residue at position 41 is preferably selected from A or P; and/or - the amino acid residue at position 89 Preferably, the amino acid residue at position 108 is selected from T, V or L; and/or - The amino acid residue at position 110 is preferably selected from Q or L; and/or - The amino acid residue at position 110 It is preferably selected from T, K or Q; and/or - The amino acid residue at position 112 is preferably selected from S, K or Q. As mentioned in these US provisional applications, which are also pending, these mutations effectively prevent or reduce the binding of so-called "pre-existing antibodies" to immunoglobulins and the compounds of the invention. For this purpose, MMP13 binding agents of the invention (such as ISVDs and/or polypeptides of the invention) may also contain (optionally in combination with such mutations) a C-terminal extension (X)n (where n is 1 to 10 , preferably 1 to 5, such as 1, 2, 3, 4 or 5 (and preferably 1 or 2, such as 1); and each X is an independently selected (preferably naturally occurring) amino acid residue , and preferably independently selected from the group consisting of: alanine (A), glycine (G), valine (V), leucine (L) or isoleucine (I) ), which further refers to those US provisional applications and WO 12/175741. In particular, MMP13 binding agents of the invention (such as ISVDs and/or polypeptides of the invention) may contain such C-terminal extensions when they form the C-terminus of proteins, polypeptides or other compounds or constructs comprising them. (See also these US provisional applications and further instructions in WO 12/175741). The MMP13 binding agent of the invention may be derived from an immunoglobulin in any suitable manner and from any suitable source, such as an immunoglobulin single variable domain, and may be, for example, a naturally occurring V_HH Sequences (i.e. from a suitable species of the family Camelidae) or synthetic or semi-synthetic amino acid sequences, including but not limited to "humanized" (as defined herein) Nanobodies or VHH sequences, "camelized" (e.g. As defined herein) immunoglobulin sequences (and in particular camelized heavy chain variable domain sequences) and Nanobodies obtained by techniques such as affinity maturation (e.g. starting from synthetic, random or naturally generated immunoglobulin sequences) , CDR grafting, veneering, combining fragments derived from different immunoglobulin sequences, PCR assembly using overlapping primers, and similar techniques for engineering immunoglobulin sequences that are well known to those skilled in the art; or any of the foregoing. Any suitable combination of the two, as further described herein. Furthermore, when immunoglobulins contain V_HH sequence, the immunoglobulin may be suitably humanized, as further described herein, to provide one or more further (partially or fully) humanized immunoglobulins of the invention. Likewise, when an immunoglobulin contains synthetic or semi-synthetic sequences (such as partially humanized sequences), then the immunoglobulin can optionally be further suitably humanized and provided with one or more further (Partially or fully) humanized immunoglobulins of the invention. "Domain antibodies", also known as "Dabs", "domain antibodies" and "dAbs" (the terms "domain antibodies" and "dAbs" are used as trademarks by the GlaxoSmithKline group of companies) have been described, for example, in EP 0368684, Ward et al. Nature 341: 544-546, 1989), Holt et al. (Tends in Biotechnology 21: 484-490, 2003) and WO 03/002609, and for example WO 04/068820, WO 06/030220, WO 06/003388 and Domantis Ltd Other published patent applications of . Domain antibodies essentially correspond to the VH or VL domains of non-camelized mammalian, specifically human 4-chain antibodies. In order to bind an epitope into a single antigen binding domain, that is, not paired with a VL or VH domain respectively, specific selection for these antigen binding properties is required, for example by using a library of human single VH or VL domain sequences. Like VHH, domain antibodies have a molecular weight of about 13 to about 16 kDa and, if derived from fully human sequences, do not require humanization for therapeutic use in, eg, humans. It should also be noted that although less preferred in the context of the present invention as they are not of mammalian origin, single variable domains may be derived from specific shark species (e.g. so-called "IgNAR domains", see e.g. WO 05/18629) . The present invention relates specifically to ISVDs, wherein the ISVDs are selected from the group consisting of VHHs, humanized VHHs and camelized VHs. The amino acid residues of the VHH domain are based on those given by Kabat et al. ("Sequence of proteins of immunological interest", US Public Health Services, NIH Bethesda, MD, Issue 91) for V_H The amino acid residue numbering of nanobodies is numbered according to the universal numbering of domains, which also applies to VHH domains from Camelidae, as in, for example, Riechmann and Muyldermans (J. Immunol. Methods 231: 25-38, 1999) As shown in Figure 2. for number V_H Methods for substitution of amino acid residues in domains are known in the art and can be applied to VHH domains in a similar manner. However, in the specification, claims, and drawings of the present invention, numbering according to Kabat as described above applies to the VHH domain, unless otherwise indicated herein. It should be noted that the V_H domains and VHH domains, the total number of amino acid residues in each CDR may vary and may not correspond to the total number of amino acid residues indicated by the Kabat numbering (i.e., one or more positions according to the Kabat numbering may not be occupied in the true sequence or the true sequence may contain more amino acid residues than Kabat numbering allows). This means that the numbering generally according to Kabat may or may not correspond to the actual numbering of the amino acid residues in the actual sequence. The total number of amino acid residues in the VH domain and VHH domain generally ranges from 110 to 120, often between 112 and 115. However, it should be noted that smaller and longer sequences may also be suitable for the purposes described herein. Regarding CDRs as they are well known in the art, there are many conventions for defining and describing CDRs of VH or VHH fragments, such as the Kabat definition (which is based on sequence variability and is most commonly used) and the Chothia definition (which is based on structural loop regions). Location). See for example the URL http://www.bioinf.org.uk/abs/. For the purposes of this specification and patent claim, CDRs are best defined based on the Abm definition (which is based on Oxford Molecular's AbM antibody modeling software) as this is considered to be the most appropriate between the Kabat and Chothia definitions. chemical compromise (refer to http://www.bioinf.org.uk/abs/). As used herein, FR1 includes the amino acid residues at positions 1 to 25, CDR1 includes the amino acid residues at positions 26 to 35, FR2 includes the amino acid residues at positions 36 to 49, and CDR2 includes Amino acid residues at positions 50 to 58, FR3 includes amino acid residues at positions 59 to 94, CDR3 includes amino acid residues at positions 95 to 102, and FR4 includes amino acid residues at positions 103 to Amino acid residue on 113. Within the meaning of the present invention, the term "immunoglobulin single variable domain" or "single variable domain" includes polypeptides derived from non-human sources, preferably Camelidae, preferably Camelidae heavy chain antibodies. They can be humanized as described herein. Furthermore, the term encompasses polypeptides derived from non-Camelid sources, such as mouse or human, which are "camelized" as described herein. Therefore, ISVDs such as domain antibodies and nanobodies (including VHH domains) can undergo humanization. In particular, a humanized ISVD such as a Nanobody (including a VHH domain) may be an ISVD as broadly defined herein, but in which at least one amino acid residue is present (and in particular in at least one of the framework residues) ), which is and/or corresponds to an ISVD of a humanized substitution (as defined herein). Potentially useful humanized substitutions can be made by comparing naturally occurring V_HH Sequences of framework regions that are closely related to one or more human V_H sequence corresponding to the framework sequence, and subsequently determined that one or more of the potentially useful humanizing substitutions (or combinations thereof) can be introduced into the V_HH sequence (in any manner known per se, as further described herein) and testable in the resulting humanized V_HH The sequence's affinity for the target, stability, ease and level of expression, and/or other desired characteristics. In this manner, one skilled in the art may, by means of a limited degree of trial and error, determine other suitable humanizing substitutions (or suitable combinations thereof) based on the disclosure herein. Furthermore, based on the foregoing, ISVD (the framework region) such as Nanobodies (including the VHH domain) can be partially or fully humanized. Another particularly preferred class of ISVDs of the present invention includes those having genes corresponding to naturally occurring V_H The amino acid sequence of the domain, but the ISVD of the amino acid sequence that has been "camelized", that is, in the naturally occurring V from the conventional 4-chain antibody_H One or more amino acid residues in the amino acid sequence of the domain appear in the V of the heavy chain antibody._HH One or more of the amino acid residues at corresponding positions in the domain are replaced. This can be done in a known manner per se apparent to those skilled in the art, for example based on the manner described herein. These "camelized" substitutions are preferably formed and/or present in V_H -V_L Interface and/or insertion at an amino acid position of a so-called camelid signature residue, as defined herein (see also eg WO 94/04678 and Davies and Riechmann (1994 and 1996)). Preferably, V is used as a starting material or starting point for the generation or design of camelized immunoglobulin single variable domains._H The sequence is preferably from mammalian V_H Sequence, better for human V_H sequence, such as V_H 3 sequence. However, it should be noted that the camelized immunoglobulin single variable domains of the present invention can be obtained in any suitable manner known per se and are therefore not strictly limited to use containing naturally occurring V_H A polypeptide obtained from a polypeptide whose domain is used as a starting material. Reference, for example, Davies and Riechmann (FEBS 339: 285-290, 1994; Biotechnol. 13: 475-479, 1995; Prot. Eng. 9: 531-537, 1996) and Riechmann and Muyldermans (J. Immunol. Methods 231: 25 -38, 1999). For example, as further explained in this article, both "humanization" and "camelization" can be achieved by providing respectively encoding naturally occurring V_HH Domain or V_H The nucleotide sequence of the domain and then changing one or more codons in the nucleotide sequence in a manner known per se, in such a way that the new nucleotide sequence encodes the "humanization" of the invention respectively. "or "camel-derived" ISVD. This nucleic acid can then be expressed in a manner known per se to provide the desired ISVD of the invention. Alternatively, naturally occurring V_HH Domain or V_H Based on the amino acid sequence of the domain, the amino acid sequence of the desired humanized or camelized ISVD of the present invention can be designed and subsequently synthesized using known peptide synthesis techniques, respectively. Also naturally occurring V_HH Domain or V_H Based on the amino acid sequence or nucleotide sequence of the domain, the nucleotide sequence encoding the desired humanized or camelized ISVD of the present invention can be designed and then re-synthesized using known nucleic acid synthesis techniques, respectively, and then The nucleic acids thus obtained can be expressed in a manner known per se so as to provide the desired ISVDs of the invention. ISVDs (such as domain antibodies and nanobodies) (including VHH domains and humanized VHH domains) can also undergo affinity maturation by introducing one or more changes in the amino acid sequence of one or more CDRs, with their respective This change results in the resulting ISVD having improved affinity for its respective antigen compared to the parent molecule. Affinity matured ISVD molecules of the present invention can be prepared by methods known in the art, for example by Marks et al. (Biotechnology 10:779-783, 1992), Barbas et al. (Proc. Nat. Acad. Sci, USA 91: 3809-3813, 1994), Shier et al. (Gene 169: 147-155, 1995), Yelton et al. (Immunol. 155: 1994-2004, 1995), Jackson et al. (J. Immunol. 154: 3310-9 , 1995), Hawkins et al. (J. Mol. Biol. 226: 889 896, 1992), Johnson and Hawkins (Affinity maturation of antibodies using phage display, Oxford University Press, 1996). Since ISVD (such as V_H ,V_L ,V_HH , domain antibodies or nanobodies) is also referred to herein as "formatting" the ISVD; and an ISVD that forms part of a polypeptide is said to be "formatted" formatted)" or in the "format" of the polypeptide. Examples of ways in which ISVDs can be formatted and examples of such formats will be apparent to those skilled in the art based on the disclosure herein; and such formatted immunoglobulin single variable domains constitute another aspect of the invention. Same look. Preferred CDRs are described in Table A-2. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein (i) CDR1 is selected from the group consisting of: (a) SEQ ID NO: 27, 28, 25, 26, 23, 29, 30, 31, 32, 33, 34, 35, 36 and 24; and (b) have 1, 2 or 3 amino acid differences from SEQ ID NOs: 27, 28, 25, 26, 23, 29, 30, 31, 32, 33, 34, 35, 36 and 24 The amino acid sequence of , 51, 38 and 39; and (d) having 1, 2 or Amino acid sequences that differ by 3 amino acids; and (iii) CDR3 is selected from the group consisting of: (e) SEQ ID NO: SEQ ID NO: 56, 107, 57, 54, 106, 55, 52, 58, 59, 60, 61, 62, 63, 64, 65, 66 and 53; and (f) with SEQ ID NO: 56, 107, 57, 54, 106, 55, 52, 58, 59, 60 , 61, 62, 63, 64, 65, 66 and 53 have amino acid sequences that differ by 1, 2, 3 or 4 amino acids. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein (i) CDR1 is selected from the group consisting of: (a) SEQ ID NO: 23; and (b) an amino acid sequence having 1 amino acid difference from SEQ ID NO: 23, Wherein at position 7, Y has been replaced by R; (ii) CDR2 is selected from the group consisting of: (c) SEQ ID NO:37; and (d) has 1, 2 or 1 with SEQ ID NO:37 Amino acid sequence with 3 amino acid differences, in which - at position 4, V has been replaced with T; - at position 5, G has been replaced with A; and/or - at position 9, N has been replaced with H; (iii) CDR3 is selected from the group consisting of: (e) SEQ ID NO: 52; and (f) an amino acid sequence having 1 amino acid difference from SEQ ID NO: 52, wherein At position 6, Y has been replaced by S. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein (i) CDR1 is SEQ ID NO:26; (ii) CDR2 is SEQ ID NO:41; and (iii) CDR3 is selected from the group consisting of: (e) SEQ ID NO:55; and (f) An amino acid sequence having 1 or 2 amino acid differences from SEQ ID NO:55, wherein - at position 8, N has been replaced by Q or S; and/or - at position 19, N has been replaced Replace with V or Q. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein (i) CDR1 is SEQ ID NO:28; (ii) CDR2 is SEQ ID NO:43; and (iii) CDR3 is selected from the group consisting of: (e) SEQ ID NO:57; and (f) An amino acid sequence having 1, 2, 3 or 4 amino acid differences from SEQ ID NO: 57, wherein - at position 10, D has been replaced by E, G, A, P, T, R, M, W or Y; - In position 16, M has been replaced by A, R, N, D, E, Q, Z, G, I, L, K, F, P, S, W, Y or V; - At position 17, D has been replaced by A, R, N, C, E, Q, Z, G, H, I, L, K, M, S, T, W, Y, or V; and/ or - In position 18, Y has been replaced by A, R, N, D, C, E, Q, Z, G, H, I, L, K, M, F, P, S, T, W or V . In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein - CDR1 is selected from the group consisting of: SEQ ID NO: 27, 28, 25, 26, 23, 29, 30, 31, 32, 33, 34, 35, 36 and 24; - CDR2 The system is selected from the group consisting of: SEQ ID NO: 42, 43, 40, 41, 37, 44, 45, 46, 47, 48, 49, 50, 51, 38 and 39; and - the CDR3 system is selected from Group consisting of: SEQ ID NO: 56, 107, 57, 54, 106, 55, 52, 58, 59, 60, 61, 62, 63, 64, 65, 66 and 53. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein the ISVD is selected from the group of ISVDs, wherein: - CDR1 is SEQ ID NO:27, CDR2 is SEQ ID NO:42, and CDR3 is SEQ ID NO:56; - CDR1 is SEQ ID NO:28 , CDR2 is SEQ ID NO:43, and CDR3 is SEQ ID NO:107; - CDR1 is SEQ ID NO:28, CDR2 is SEQ ID NO:43, and CDR3 is SEQ ID NO:57; - CDR1 is SEQ ID NO :25, CDR2 is SEQ ID NO:40, and CDR3 is SEQ ID NO:54; - CDR1 is SEQ ID NO:26, CDR2 is SEQ ID NO:41, and CDR3 is SEQ ID NO:106; - CDR1 is SEQ ID NO:26, CDR2 is SEQ ID NO:41, and CDR3 is SEQ ID NO:55; - CDR1 is SEQ ID NO:23, CDR2 is SEQ ID NO:37, and CDR3 is SEQ ID NO:52; - CDR1 is SEQ ID NO:26, CDR2 is SEQ ID NO:48, and CDR3 is SEQ ID NO:62; - CDR1 is SEQ ID NO:26, CDR2 is SEQ ID NO:41, and CDR3 is SEQ ID NO:63; - CDR1 is SEQ ID NO:29, CDR2 is SEQ ID NO:44, and CDR3 is SEQ ID NO:58; - CDR1 is SEQ ID NO:30, CDR2 is SEQ ID NO:45, and CDR3 is SEQ ID NO: 58; - CDR1 is SEQ ID NO:31, CDR2 is SEQ ID NO:46, and CDR3 is SEQ ID NO:59; - CDR1 is SEQ ID NO:32, CDR2 is SEQ ID NO:47, and CDR3 is SEQ ID NO:60; - CDR1 is SEQ ID NO:33, CDR2 is SEQ ID NO:41, and CDR3 is SEQ ID NO:61; - CDR1 is SEQ ID NO:34, CDR2 is SEQ ID NO:49, and CDR3 is SEQ ID NO:64; - CDR1 is SEQ ID NO:35, CDR2 is SEQ ID NO:50, and CDR3 is SEQ ID NO:65; - CDR1 is SEQ ID NO:36, CDR2 is SEQ ID NO:51, and CDR3 is SEQ ID NO:66; - CDR1 is SEQ ID NO:23, CDR2 is SEQ ID NO:39, and CDR3 is SEQ ID NO:53; and - CDR1 is SEQ ID NO:24, CDR2 is SEQ ID NO: 38, and CDR3 is SEQ ID NO:52. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein CDR1 is SEQ ID NO:27, CDR2 is SEQ ID NO:42, and CDR3 is SEQ ID NO:56. In particular, the present invention relates to an ISVD as described herein, wherein the ISVD specifically binds to MMP13 and consists essentially of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively) Composed of, wherein the ISVD is selected from the group consisting of: SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10, 1, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 2, 3, 4, 5, 6, 7 and 8. It should be understood, without limitation, that the immunoglobulin single variable domains of the present invention can be used as "building blocks" for preparing polypeptides, which may optionally contain one or more other immunoglobulin single variable domains that can be used as building blocks. (i.e., against the same or another epitope on MMP13 and/or against one or more other antigens, proteins or targets besides MMP13). Polypeptides The polypeptides of the invention comprise at least one ISVD that binds to a MMP, preferably MMP13, such as two ISVDs that bind to MMP13, and preferably also comprise at least one ISVD that binds aggrecan, more preferably two. An ISVD that binds to aggrecan. In the polypeptide of the invention, ISVD can be linked directly or via a linker. Even more preferably the polypeptides of the invention comprise a C-terminal extension. As detailed below, C-terminal extension substantially prevents/removes binding of pre-existing antibodies/factors in most samples from human individuals/patients. The C-terminal extension is present at the C-terminus of the last amino acid residue (often a serine residue) of the last (mostly C-terminally positioned) ISVD. As further detailed in this article, ISVD can be derived from V_HH ,V_H or V_L domains, however the ISVDs are selected such that they do not form V in the polypeptides of the invention._H with V_L Complementary pairs of domains. Nanobodies, V_HH and humanized V_HH are unusual because they are derived from native camelid antibodies that do not have light chains and the fact that these domains are unable to associate with camelid light chains to form complementary V_HH with V_L right. Therefore, the polypeptides of the invention do not comprise complementary ISVDs and/or form complementary ISVD pairs, such as complementary V_H /V_L right. Polypeptides or constructs that generally contain or consist essentially of a single building unit (such as a single ISVD or a single Nanobody) are referred to herein as "monovalent" polypeptides and "monovalent constructs" respectively. Polypeptides or constructs containing two or more building blocks (such as ISVD) are also referred to herein as "multivalent" polypeptides or constructs and the building blocks/ISVDs present in such polypeptides or constructs are also referred to herein as It is in "multi-price format". For example, a "bivalent" polypeptide may comprise two ISVDs, optionally linked via a linker sequence, while a "trivalent" polypeptide may comprise three ISVDs, optionally linked via two linker sequences, and a "tetravalent" polypeptide may Contains four ISVDs optionally connected via three linker subsequences; etc. In a multivalent polypeptide, two or more ISVDs may be the same or different and may be directed against the same antigen or epitope (e.g., against the same part or epitope or against different parts or epitopes) or may Another alternative is to target different antigens or antigenic determinants, or any suitable combination thereof. Polypeptides and constructs containing at least two building blocks (such as ISVD), wherein at least one building block is directed against a first antigen (i.e., MMP13) and at least one building block is directed against a second antigen (i.e., different from MMP13) ) are also referred to as "multispecific" polypeptides and constructs, and the building blocks (such as ISVD) present in such polypeptides and constructs are also referred to herein as being in a "multispecific format." Thus, for example, a "bispecific" polypeptide of the invention is a polypeptide comprising at least one ISVD directed against a first antigen (i.e., MMP13) and at least one additional ISVD directed against a second antigen (i.e., different from MMP13) , and the "trispecific" polypeptide of the present invention is one that includes at least one ISVD targeting the first antigen (i.e., MMP13), at least one additional ISVD targeting the second antigen (i.e., different from MMP13), and at least one ISVD targeting the second antigen (i.e., different from MMP13). Polypeptides of additional ISVDs directed against a third antigen (i.e., different from both MMP13 and the second antigen); etc. In one aspect, the invention relates to polypeptides comprising two or more ISVDs that specifically bind to MMP13, wherein a) at least the "first" ISVD is related to the first epitope, epitope region, portion, Domain, subunit or conformation specifically binds; preferably the "first" ISVD that specifically binds to MMP13 is selected from the group consisting of: SEQ ID NO: 111, 11, 110, 10, 112 , 12, 109, 9, 13, 14, 15, 16, 17, 18, 19, 20, 21 and 22; and among them b) at least the "second" ISVD is related to the second antigenic determinant and antigenic determinant of MMP13 , a portion, domain, subunit or configuration that specifically binds to the first epitope, epitope, region, portion, domain, subunit or configuration, respectively, that preferably specifically binds to MMP13 Two "ISVDs" were selected from the group consisting of: SEQ ID NO: 1, 2, 3, 4, 5, 6, 7 and 8. In one aspect, the present invention relates to polypeptides comprising two or more ISVDs that specifically bind to MMP13, which are selected from the group consisting of: SEQ ID NO: 160 to 165, preferably SEQ ID NO :160 (refer to Table A-3). "Multiparatope" polypeptides and "multiparatope" constructs (such as "biparatope" polypeptides or constructs and "triparatope" polypeptides or constructs) comprise or consist essentially of two or more paratopes each having different paratopes. Composed of more building units. Accordingly, the ISVDs of the invention that bind to MMP13 may be in a substantially isolated form (as defined herein), or they may form part of a construct or polypeptide, which may comprise or consist essentially of one or more MMP13-binding ISVDs. ISVD consists of, and may optionally additionally contain, one or more additional amino acid sequences (all optionally linked via one or more suitable linkers). The invention relates to a polypeptide or construct comprising or consisting essentially of at least one ISVD according to the invention (such as one or more ISVDs of the invention) (or a suitable fragment thereof) that binds to MMP13. One or more ISVDs of the invention may be used as building blocks in such polypeptides or constructs to provide monovalent, multivalent or multiparatopic polypeptides or constructs of the invention, respectively, all as described herein. The invention therefore also relates to polypeptides that are monovalent constructs, which comprise or consist essentially of a monovalent polypeptide or ISVD of the invention. The invention therefore also relates to polypeptides or constructs, respectively, multivalent polypeptides or multivalent constructs, such as bivalent or trivalent polypeptides or constructs, which comprise or consist essentially of two or more ISVDs of the invention (regarding Multivalent and multispecific polypeptides containing one or more VHH domains and their preparation are also referred to, for example, Conrath et al. (J. Biol. Chem. 276: 7346-7350, 2001), and for example, WO 96/34103, WO 99/23221 and WO 2010/115998). In one aspect, in its simplest form, a multivalent polypeptide or construct of the invention is a bivalent polypeptide or construct of the invention comprising a first ISVD (such as a Nanobody) directed against MMP13 and directed to the same second ISVD (such as a Nanobody) directed against MMP13, wherein the first and the second ISVD (such as a Nanobody) are optionally linked via a linker sequence (as defined herein). In its simplest form, a multivalent polypeptide or construct of the invention may be a trivalent polypeptide or construct of the invention, comprising a first ISVD (such as a Nanobody) directed against MMP13, The same second ISVD (such as a nanobody) and the same third ISVD (such as a nanobody) directed against MMP13, wherein the first, second and third ISVD (such as a nanobody) They may optionally be connected via one or more, and in particular two, linker subsequences. In one aspect, the invention relates to a polypeptide or construct comprising or consisting essentially of at least two ISVDs according to the invention (such as 2, 3 or 4 ISVDs) (or suitable fragments thereof) that bind to MMP13 composition. Two or more ISVDs can optionally be linked via one or more peptide linkers. In another aspect, a multivalent polypeptide or construct of the invention may be a bispecific polypeptide or construct of the invention, comprising a first ISVD (such as a Nanobody) directed against MMP13 and a second ISVD directed against A second ISVD (such as a Nanobody) of an antigen (such as aggrecan), wherein the first and second ISVD (such as a Nanobody) are optionally linked via a linker sequence (as defined herein) ; And the multivalent polypeptide or construct of the present invention can also be the trispecific polypeptide or construct of the present invention, which includes a first ISVD directed against MMP13 (such as a nanobody), a second antigen directed against (such as aggregation a second ISVD (such as a nanobody) targeting a proteoglycan) and a third ISVD (such as a nanobody) directed against a third antigen, wherein the first, second and third ISVD (such as a nanobody) Antibodies) can optionally be linked via one or more, and in particular two, linker sequences. The invention further relates to multivalent polypeptides comprising or (essentially) consisting of at least one ISVD (or a suitable fragment thereof) that binds MMP13, preferably human MMP13, and at least one additional ISVD such as aggrecan-binding of ISVD). Particularly preferred trivalent, bispecific polypeptides or constructs according to the invention are those set forth in the Examples described herein and Table A-3. In a preferred aspect, the polypeptide or construct of the invention comprises or consists essentially of at least two ISVDs, wherein the at least two ISVDs may be the same or different, but at least one of the ISVDs is directed against MMP13. Two or more ISVDs present in a multivalent polypeptide or construct of the invention may be composed of light chain variable domain sequences (e.g., V_L - sequence) or heavy chain variable domain sequence (e.g. V_H - sequences); they may be composed of heavy chain variable domain sequences derived from conventional quadruplex antibodies or heavy chain variable domain sequences derived from heavy chain antibodies. In a preferred aspect, they are domain antibodies (or peptides suitable for use as domain antibodies), single domain antibodies (or peptides suitable for use as single domain antibodies), "dAb" ( or peptides suitable for use as dAb), Nanobody® (including but not limited to V_HH ), humanized V_HH Sequence, camel-derived V_H Sequence or V_HH composed of sequences. Two or more immunoglobulin single variable domains may be composed of partially or fully humanized Nanobodies or partially or fully humanized VHHs. In aspects of the invention, the first ISVD and the second ISVD present in the multiparatope (preferably biparatope or triparatope) polypeptide or construct of the invention do not (cross) compete with each other. A combination of MMP13s, and indeed belong to different families. Accordingly, the present invention relates to polypeptides or constructs comprising two or more ISVDs, preferably biparatopes, where each ISVD belongs to a different family. In one aspect, the first ISVD of the multiparatope (preferably biparatope) polypeptide or construct of the invention does not cross-block the multiparatope (preferably biparatope) polypeptide of the invention Or the second ISVD of the construct binds to MMP13 and/or the first ISVD is not cross-blocked by the second ISVD from binding to MMP13. In another aspect, the first ISVD cross-blocking multiparatope (preferably biparatope) polypeptide or construct of the invention blocks the multiparatope (preferably biparatope) polypeptide of the invention or The second ISVD of the construct binds to MMP13 and/or the first ISVD is cross-blocked by the second ISVD from binding to MMP13. In a particularly preferred aspect, a polypeptide or construct of the invention comprises or consists essentially of three or more ISVDs, at least two of which are directed against MMP13. It should be understood that the at least two ISVDs targeting MMP13 can be the same or different, can target the same epitope or different epitopes of MMP13, and can belong to the same epitope bin or different epitopes. Bins and/or may bind to the same or different MMP13 domains. In a preferred aspect, the polypeptide or construct of the invention comprises or essentially consists of at least two ISVDs that bind to MMP13, wherein the at least two ISVDs may be the same or different, and are independently selected from the following: Group consisting of SEQ ID NO: 111, 11, 110, 10, 112, 12, 109, 9, 13, 14, 15, 16, 17, 18, 19, 20, 21 and 22 and SEQ ID NO: 1, 2, 3, 4, 5, 6, 7 and 8, preferably the at least two ISVDs are independently selected from the group consisting of: SEQ ID NO: 111, 11, 110, 10 , 112, 12, 109, 9 and 1, and/or at least one ISVD is selected from the group consisting of: SEQ ID NO: 111, 11, 110, 10, 112, 12, 109, 9, 13, 14 , 15, 16, 17, 18, 19, 20, 21 and 22 and at least one ISVD are selected from the group consisting of: SEQ ID NO: 1, 2, 3, 4, 5, 6, 7 and 8. In another aspect, the invention relates to polypeptides or constructs comprising two or more multiparatopic (preferably biparatopic) ISVDs directed against MMP13 that bind to the same epitope, e.g. Combined with any of the following: SEQ ID NO: 111, 11, 110, 10, 112, 12, 109, 9, 13, 14, 15, 16, 17, 18, 19, 20, 21 and 22 or the following Combining any of: SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7 and 8. In another aspect, the invention relates to a polypeptide as described herein, wherein the polypeptide has at least 80%, 90%, 95% or 100% (more preferably at least 95%, and most preferably 100%) sequence identity: SEQ ID NO: 160 to 165 (i.e. 160, 161, 162, 163, 164 or 165) and 176 to 192 (i.e. 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 or 192), preferably SEQ ID NO: 192. Sparing This technology is needed for more effective treatment of diseases affecting joint cartilage, such as osteoarthritis. Even when administered intra-articularly, most drugs used to treat affected cartilage do not have sufficient residence time. The inventors speculate that the efficacy of therapeutic agents (such as the constructs, polypeptides and ISVDs of the present invention) may be achieved by combining the therapeutic agent with a device that "anchors" the drug in the joint and subsequently increases drug retention, but should not destroy the therapeutic agent. Part of the efficacy is coupled and significantly increased (this part is also represented by "anchored protein cartilage" or "CAP" in this article). This anchoring concept not only increases drug efficacy by reducing toxicity and side effects, but also increases operational specificity for diseased joints, thereby expanding the number of potentially useful drugs. The final format of the molecule expected for clinical use contains one or two building blocks that bind to MMP13 (such as ISVD) and one or more building blocks that have this mode of action retention (such as ISVD), and possibly more. In the Examples section, it is demonstrated that these formats retain both MMP13 binding and therapeutic effects, such as inhibitory activity and retention properties. One or more building blocks with a retained mode of action (e.g., ISVD) can be any building block ("CAP building block") with retained effects in diseases involving MMP13, such as arthritic diseases, osteoarthritis, spondylosis Insufficiency, degenerative lumbar disc disease, degenerative joint disease, rheumatoid arthritis, osteochondritis dissecans, aggrecanopathies and malignant tumor metastasis. "CAP building blocks" are used to guide, anchor, and/or retain other, e.g., therapeutic building blocks (such as ISVDs that bind to MMP13) in a desired location (such as in a joint), where the other, e.g., therapeutic, e.g. The building blocks exert their effects, such as binding and/or inhibiting MMP13 activity. The inventors additionally speculate that aggrecan binding agents, such as ISVD that binds aggrecan, may serve as such anchors, although aggrecan is highly glycated and degraded in various disorders affecting articular cartilage. Furthermore, given the cost and large-scale testing required in various animal models before drugs can enter the clinic, such aggrecan-binding agents should preferably have broad species cross-reactivity, e.g., aggrecan-binding agents should be compatible with Aggrecan binding in various species. The inventors were able to use a variety of ingenious immunological, screening and characterization methods to identify a variety of aggrecan binders with excellent selectivity, stability and specificity properties that result in prolonged retention and activity in joints. In one aspect, the invention relates to a method of reducing and/or inhibiting the outflow of a composition, polypeptide or construct from a joint, wherein the method comprises administering to a subject in need thereof a pharmaceutically active amount of at least one polypeptide according to the invention, A construct according to the invention or a composition according to the invention. In the present invention, the term "reducing and/or inhibiting outflow" means reducing and/or inhibiting the outward flow of a composition, polypeptide or construct from the inside of a joint to the outside. Preferably efflux compared to the efflux of the aforementioned composition, polypeptide or construct in the joint under the same conditions but in the absence of an aggrecan-binding agent of the invention (e.g., an ISVD that binds aggrecan). Reduction and/or inhibition by at least 10%, such as at least 20%, 30%, 40% or 50% or even higher, such as at least 60%, 70%, 80%, 90% or even 100%. Next are diseases in which MMP13 is involved, such as arthritic diseases, osteoarthritis, spondylodysplasia, degenerative lumbar disc disease, degenerative joint disease, rheumatoid arthritis, osteochondritis dissecans, aggrecan lesions and malignant tumor metastasis, it is expected that the aggrecan binding agent of the present invention can also be used for various other diseases affecting cartilage, such as joint lesions and chondrodystrophies, arthritic diseases (such as osteoarthritis, rheumatoid arthritis, gout arthritis, psoriatic arthritis, traumatic rupture or detachment), achondroplasia, costochondritis, aplasia of the spine, disc herniation, degenerative lumbar disc disease, degenerative joint disease, and relapsing polychondritis ( Often referred to as "aggrecan-related diseases" in this article). CAP building blocks (e.g., ISVD bound to aggrecan) preferably bind to cartilaginous tissue (such as cartilage and/or meniscus). In preferred aspects, the CAP building blocks are cross-reactive to other species and are compatible with human aggrecan (SEQ ID NO: 105), dog aggrecan, bovine aggrecan, rat aggrecan, Specific binding to one or more of the sugar, porcine aggrecan, mouse aggrecan, rabbit aggrecan, cynomolgus aggrecan and/or rhesus monkey aggrecan. Relevant structural information for aggrecan can be found, for example, in the (UniProt) accession numbers described in Table 2 below. Preferred CAP building blocks are ISVDs that bind to aggrecan, preferably human aggrecan, preferably represented by SEQ ID NO: 105, as described in Table B. The invention therefore relates to polypeptides or constructs according to the invention, which additionally comprise at least one CAP building block. The invention therefore relates to a polypeptide or construct according to the invention, which further comprises at least one ISVD that specifically binds aggrecan, such as shown in Table E, and preferably selected from the group consisting of SEQ ID NO: 166 to 168 Represented by ISVD. In particular, the present invention relates to an ISVD that specifically binds to aggrecan, wherein the ISVD essentially consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), wherein the ISVD is selected from the group of ISVDs, wherein (a) CDR1 is SEQ ID NO:169, CDR2 is SEQ ID NO:170; and CDR3 is SEQ ID NO:171; and (b) CDR1 is SEQ ID NO: 172, CDR2 is SEQ ID NO:173, and CDR3 is SEQ ID NO:174. In one aspect, the invention relates to a polypeptide as described herein, comprising at least 2 ISVDs that specifically bind aggrecan. In one aspect, the invention relates to a polypeptide as described herein comprising at least 2 ISVDs that specifically bind to aggrecan, wherein the at least 2 ISVDs that specifically bind to aggrecan may be the same or different. In one aspect, the invention relates to a polypeptide as described herein comprising at least 2 ISVDs that specifically bind to aggrecan, wherein the at least 2 ISVDs that specifically bind to aggrecan are independently Selected from the group consisting of: SEQ ID NO: 166 to 168. In one aspect, the invention relates to a polypeptide as described herein, comprising at least 2 ISVDs that specifically bind to aggrecan, wherein the at least 2 ISVDs that specifically bind to aggrecan are represented by SEQ. ID NO:166 to 168 represents. In one aspect, the invention relates to a polypeptide as described herein, comprising an ISVD that specifically binds aggrecan, wherein the ISVD that specifically binds aggrecan is with human aggrecan [SEQ ID NO:105] Specific binding. In one aspect, the invention relates to polypeptides as described herein, wherein the ISVD that specifically binds aggrecan is with human aggrecan (SEQ ID NO: 105), dog aggrecan, bovine aggrecan, Aggrecan, rat aggrecan, porcine aggrecan, mouse aggrecan, rabbit aggrecan, macaque aggrecan and/or rhesus monkey aggrecan specific combine. In one aspect, the invention relates to polypeptides as described herein, wherein the ISVD that specifically binds to aggrecan preferably binds to cartilaginous tissue (such as cartilage and/or meniscus). It is understood that the ISVDs, polypeptides and constructs of the invention are preferably stable. The stability of a polypeptide, construct or ISVD of the invention can be measured by conventional assays known to those skilled in the art. Typical assays include, without limitation, assays in which the activity of such polypeptides, constructs or ISVDs is determined, followed by incubation in synovial fluid for a desired period, and subsequent activity is determined, such as those detailed in the Examples paragraph. In one aspect, the invention relates to an ISVD, polypeptide or construct of the invention having stability in synovial fluid (SF) at 37°C for at least 7 days, such as at least 14 days, 21 days, 1 month , 2 months or even 3 months. In one aspect, the invention relates to ISVDs, polypeptides or constructs of the invention that penetrate at least 5 microns into cartilage, such as at least 10 microns, 20 microns, 30 microns, 40 microns, 50 microns or even more. The desired activity of a therapeutic building block (eg, an ISVD binding to MMP13 in a multivalent polypeptide or construct of the invention) can be measured by conventional assays known to those skilled in the art. Typical assays include (non-limiting) GAG release assays, as detailed in the Examples paragraph. Relative affinity can depend on the position of the ISVD within the polypeptide. It will be understood that the order (orientation) of the ISVDs in the polypeptides of the invention can be selected according to the needs of one skilled in the art. The order of individual ISVDs and whether the polypeptide contains a linker is a matter of design choice. Some orientations, with or without linkers, may provide better binding characteristics than other orientations. For example, the order of the first ISVD (e.g., ISVD 1) and the second ISVD (e.g., ISVD 2) in the polypeptide of the invention can be (from N-terminus to C-terminus): (i) ISVD 1 (e.g., ISVD 1) Nanobody 1)-[linker]-ISVD 2 (e.g. Nanobody 2)-[C-terminal extension]; or (ii) ISVD 2 (e.g. Nanobody 2)-[linker]-ISVD 1( For example, Nanobody 1)-[C-terminal extension]; (the part between the square brackets (i.e., the linker and the C-terminal extension) is optional). The present invention covers all orientations. Polypeptides containing ISVD orientations that provide desired binding characteristics can be readily identified by routine screening, such as exemplified in the Examples paragraph. The preferred sequence is from N-terminus to C-terminus: ISVD-[linker] that binds to MMP13-ISVD-[C-terminal extension] that binds to aggrecan, with the part between square brackets For random. A better sequence is from N-terminal to C-terminal: ISVD-[linker] that binds to MMP13-ISVD-[linker] that binds to aggrecan-ISVD-[C that binds to aggrecan -end extension], where the part between square brackets is optional. See, for example, Table F. Half-life In certain aspects of the invention, a construct or polypeptide of the invention may have a moiety that confers an increased half-life compared to a corresponding construct or polypeptide of the invention without such moiety. Some preferred but non-limiting examples of these constructs and polypeptides of the invention will be apparent to those skilled in the art based on the further disclosure herein, and include, for example, the ISVD of the invention that is chemically modified to increase its half-life or A polypeptide (e.g. by PEGylation); an MMP13 binding agent of the invention (such as an ISVD and/or polypeptide of the invention) comprising at least one additional binding site for binding to a serum protein (such as serum albumin); or comprising a Polypeptides of the invention that are linked to at least one ISVD of the invention linked to at least one portion (and in particular at least one amino acid sequence) that increases the half-life of an amino acid sequence of the invention. Polypeptides of the invention comprising such half-life extending portions or ISVDs Examples of constructs (such as polypeptides of the invention) will be apparent to those skilled in the art based on the further disclosure herein; and examples include, but are not limited to, polypeptides in which one or more ISVDs of the invention are combined with one or A plurality of serum proteins or fragments thereof (such as (human) serum albumin or suitable fragments thereof) or polypeptides suitably linked to one or more binding units capable of binding to a serum protein (such as domain antibodies, suitable for use as domain antibodies An immunoglobulin single variable domain, a single domain antibody, an immunoglobulin single variable domain suitable for use as a single domain antibody, a dAb, an immunoglobulin single variable domain suitable for use as a dAb, or may be combined with a serum protein such as Serum albumin, such as human serum albumin), serum immunoglobulin (such as IgG) or transferrin-binding Nanobodies; see further description and references mentioned herein); wherein the amino acids of the present invention A polypeptide whose sequence is linked to an Fc portion (such as human Fc), or a suitable portion or fragment thereof; or wherein one or more immunoglobulin single variable domains of the invention are linked to one or more small molecules capable of binding to a serum protein. Proteins or peptides suitably linked polypeptides, such as those described in WO 91/01743, WO 01/45746, WO 02/076489, WO2008/068280, WO2009/127691 and PCT/EP2011/051559. In one state In this manner, the invention provides a construct or polypeptide of the invention, wherein the construct or the polypeptide additionally comprises a serum protein binding moiety or a serum protein. Preferably the serum protein binding moiety is combined with serum albumin (such as human serum albumin ) binds. In one aspect, the invention relates to a polypeptide as described herein comprising an ISVD that binds to serum albumin. Constructs and polypeptides of the invention that generally have an increased half-life preferably have a higher half-life than the corresponding The constructs and polypeptides themselves (i.e. without portions conferring increased half-life) have a half-life that is at least 1.5 times greater, preferably at least 2 times greater, such as at least 5 times, such as at least 10 times or more than 20 times greater. For example, having an increased half-life The construct or polypeptide of the invention may, for example, have an increased half-life of more than 1 hour, preferably more than 2 hours, compared to the corresponding construct or polypeptide of the invention itself (i.e. without a moiety conferring increased half-life), More preferably a half-life of more than 6 hours, such as more than 12 hours or even more than 24, 48 or 72 hours. In a preferred but non-limiting aspect of the invention, the constructs of the invention and the polypeptides of the invention have properties similar to those of the corresponding constructs and polypeptides of the invention themselves (i.e. without the moiety conferring increased half-life), for example in humans. A serum half-life of more than 1 hour, preferably more than 2 hours, more preferably more than 6 hours, such as more than 12 hours or even more than 24, 48 or 72 hours. In another preferred but non-limiting aspect of the invention, the constructs of the invention (such as the polypeptides of the invention) exhibit in humans at least about 12 hours, preferably at least 24 hours, more preferably at least 48 hours. hours, and even more preferably a serum half-life of at least 72 hours or longer. For example, a construct or polypeptide of the invention may have at least 5 days (such as about 5 to 10 days), preferably at least 9 days (such as about 9 to 14 days), more preferably at least about 10 days (such as about 10 to 14 days). 15 days), or at least about 11 days (such as about 11 to 16 days), more preferably at least about 12 days (such as about 12 to 18 days or more), or more than 14 days (such as about 14 to 19 days) half life. In a particularly preferred but non-limiting aspect of the invention, the invention provides constructs of the invention and polypeptides of the invention, which comprise in addition to one or more building blocks binding to MMP13 and possibly one or more binding proteins to aggregation proteins. In addition to the glycan-binding CAP building blocks (eg, ISVD), at least one building block that binds to serum albumin, such as an ISVD that binds to serum albumin (such as human serum albumin), is also included, as described herein. The ISVD that preferably binds to serum albumin includes or consists essentially of 4 framework regions (FR1 to FR4 respectively) and 3 complementarity determining regions (CDR1 to CDR3 respectively), wherein CDR1 is SFGMS and CDR2 is SISGSGSDTLYADSVKG, and CDR3 is GGSLSR. The ISVD that preferably binds to human serum albumin is selected from the group consisting of: Alb8, Alb23, Alb129, Alb132, Alb11, Alb11(S112K)-A, Alb82, Alb82-A, Alb82-AA, Alb82 -AAA, Alb82-G, Alb82-GG, Alb82-GGG, Alb92, Alb135 or Alb223 (refer to Table D). In one embodiment, the invention relates to a construct (such as a polypeptide) of the invention comprising a serum protein binding moiety, wherein the serum protein binding moiety is a non-antibody based polypeptide. Other Moieties In one aspect, the invention relates to constructs as described herein, which comprise one or more other groups, residues, moieties or binding units. The one or more other groups, residues, moieties or binding units are preferably selected from the group consisting of polyethylene glycol molecules, serum proteins or fragments thereof, binding units capable of binding to serum proteins, Fc Parts and small proteins or peptides that can bind to serum proteins, additional amino acid residues, tags or other functional parts, such as toxins, markers, radiochemicals, etc. In the embodiments mentioned below, the invention relates to constructs of the invention (such as polypeptides) comprising a moiety conferring half-life extension, wherein the moiety is PEG. Therefore, the invention also relates to constructs or polypeptides of the invention comprising PEG. Additional amino acid residues may or may not alter, alter or otherwise affect other (biological) properties of the polypeptides of the invention and may or may not add additional functionality to the polypeptides of the invention. For example, such amino acid residues: a) may contain an N-terminal Met residue, for example due to expression in a heterologous host cell or host organism. b) A signal sequence or leader sequence may be formed which directs the secretion of the polypeptide from the host cell upon synthesis (e.g. to provide a pre-, pro- or prepro-form) of the polypeptide of the invention, which is Depends on the host cell used to express the polypeptide of the invention). Suitable secretory leader peptides will be known to those skilled in the art and may be described further herein. Such leader sequences are often linked to the N-terminus of the polypeptide, although the invention in its broadest sense is not so limited; c) can form a "tag", such as an amino acid sequence or residue that allows or accelerates purification of the polypeptide, for example using a guide Affinity technology for that sequence or residue. The sequence or residues can then be removed (eg by chemical or enzymatic cleavage) to provide a polypeptide (for this purpose, the tag can optionally be linked to an amino acid sequence or polypeptide sequence via a cleavable linker sequence or Contains cleavable motifs). Some preferred but non-limiting examples of such residues are multiple histidine residues, glutathione residues, myc-tags (such as AAAEQKLISEEDLNGAA (SEQ ID NO: 175)), MYC-HIS-tags (SEQ ID NO:123) or FLAG-HIS6-tag (SEQ ID NO:124) (see Table B); d) can be one or more amine groups that have been functionalized and/or can serve as sites for attachment of functional groups acid residue. Suitable amino acid residues and functional groups will be apparent to those skilled in the art and include, but are not limited to, those mentioned herein with respect to derivatives of the polypeptides of the invention. The invention also encompasses constructs and/or polypeptides comprising the ISVD of the invention and additionally comprising other functional moieties (eg, toxins, markers, radiochemicals, etc.). Other groups, residues, moieties or binding units may be, for example, chemical groups, residues, moieties which themselves may or may not have biological and/or pharmacological activity. For example, and without limitation, such groups may be linked to one or more ISVDs or polypeptides of the invention to provide "derivatives" of the polypeptides or constructs of the invention. Therefore, the invention in its broadest sense also includes constructs and/or polypeptides that are derivatives of the constructs and/or polypeptides of the invention. Such derivatives generally modify the constructs and/or polypeptides of the invention and/or form one of the amino acid residues forming the polypeptides of the invention, and in particular chemically and/or biologically (e.g., enzymatically). or more. Examples of such modifications, as well as the amino acid residues within the polypeptide sequence that can be modified in this manner (i.e., on the protein backbone, but preferably on the side chains), methods and techniques that can be used to introduce such modifications , and examples of possible uses and advantages of such modifications will be apparent to those skilled in the art (see also Zangi et al., Nat Biotechnol 31(10):898-907, 2013). For example, such modifications may comprise the introduction (eg by covalent linkage or in any other suitable manner) of one or more (functional) groups, residues or moieties into or on the polypeptide of the invention, and in particular One or more functional groups, residues or moieties that confer one or more desired properties or functionality are introduced into the constructs and/or polypeptides of the invention. Examples of such functional groups will be apparent to those skilled in the art. For example, such modifications may include the introduction (e.g., covalently linked or in any other suitable manner) of one or more functional moieties that increase the half-life, solubility and/or absorption of the construct or polypeptide of the invention, decrease The immunogenicity and/or toxicity of the construct or polypeptide of the invention, eliminate or reduce any undesirable side effects of the construct or polypeptide of the invention, and/or confer other advantageous properties to the construct or polypeptide of the invention and/or or reduction of undesirable characteristics; or any combination of two or more of the foregoing. Examples of such functional components and the techniques incorporating them will be apparent to those skilled in the art, and may generally include all functional components and techniques mentioned in the general background cited above, as well as for Functional moieties and techniques known per se for modifying pharmaceutical proteins and particularly for modifying antibodies or antibody fragments (including ScFv and single domain antibodies), see for example Remington (Pharmaceutical Sciences, 16th ed., Mack Publishing Co., Easton, PA , 1980). Such functional moieties may, for example, be linked directly (eg, covalently) to the polypeptide of the invention or optionally via a suitable linker or spacer, as will be apparent to those skilled in the art. A specific example are derivatives of the polypeptide or construct of the invention, wherein the polypeptide or construct of the invention has been chemically modified to increase its half-life (eg by means of PEGylation). This is one of the most widely used techniques to increase the half-life and/or reduce the immunogenicity of pharmaceutical proteins and involves attachment of suitable pharmacologically acceptable polymers such as poly(ethylene glycol) (PEG) or its derivatives substances (such as methoxypoly(ethylene glycol) or mPEG). Generally any suitable pegylation format may be used, such as that used in the art for pegylation of antibodies and antibody fragments including but not limited to (single) domain antibodies and ScFv; see e.g. Chapman (Nat. Biotechnol .54: 531-545, 2002), Veronese and Harris (Adv.Drug Deliv.Rev.54: 453-456, 2003), Harris and Chess (Nat.Rev.Drug.Discov.2: 214-221, 2003) and WO 04/060965. Various reagents for protein pegylation are also commercially available, for example from Nektar Therapeutics, USA. Preferably site-directed PEGylation is used, in particular via cysteine residues (see e.g. Yang et al. (Protein Engineering 16: 761-770, 2003)). For example, for this purpose PEG can be attached to Naturally occurring cysteine residues in polypeptides of the invention, constructs or polypeptides of the invention may be modified to appropriately introduce one or more cysteine residues for attachment of PEG, or may include The amino acid sequence of one or more cysteine residues of PEG can be fused to the N-terminus and/or C-terminus of the construct or polypeptide of the invention, all using methods known per se to those skilled in the art. Protein engineering technology. PEG preferably used in the constructs and polypeptides of the invention has a molecular weight of more than 5000, such as more than 10,000 and less than 200,000, such as less than 100,000; for example, in the range of 20,000 to 80,000. Additionally, generally less preferred modifications include N-linked or O-linked glycation, often as part of co-translational and/or post-translational modifications, depending on the host cell used to express the polypeptide of the invention. Yet another modification may include the introduction of one or more detectable labels or other signal generating groups or moieties, depending on the intended use of the polypeptide or construct of the invention. Suitable labels and techniques for attaching, using and detecting them will be apparent to those skilled in the art, and include, but are not limited to, fluorescent labels (such as fluorescein, isothiocyanate, rose bengal, Phycoerythrin, phycocyanin, allophycocyanin, o-dialdehyde benzoic acid and fluorocarmine) and fluorescent metals (such as¹⁵² Eu or other metals from the lanthanide series), phosphorescent, chemiluminescent or bioluminescent markers (such as luminal, isoluminol, theromatic acridinium esters, imidazole, Acridinium salts, oxalates, dioxetanes or GFP and their analogs), radioactive isotopes such as³ H.¹²⁵ I.³² P.³⁵ S.¹⁴ C.⁵¹ Cr,³⁶ Cl.⁵⁷ Co.,⁵⁸ Co.,⁵⁹ Fe and⁷⁵ Se), metals, metal chelates or metal cations (e.g. metal cations such as^99m Tc,¹²³ I.¹¹¹ In,¹³¹ I.⁹⁷ Ru,⁶⁷ Cu,⁶⁷ Ga and⁶⁸ Ga or other metals or metal cations particularly suitable for use in in vivo, in vitro or on-site diagnostics and imaging, such as (¹⁵⁷ Gd,⁵⁵ Mn,¹⁶² Dy,⁵² Cr and⁵⁶ Fe)), as well as chromophores and enzymes (such as malate dehydrogenase, staphylococcal nuclease, delta-V-steroid isomerase, yeast alcohol dehydrogenase, α-glycerophosphate dehydrogenase, triosephosphate isomerase Constructase, biotin avidin peroxidase, wasabi peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, β-galactosidase, ribonuclease, urease, catalase, glucose -VI-phosphate dehydrogenase, glucoamylase and acetylcholinesterase). Other suitable labels will be apparent to those skilled in the art, and include, for example, moieties detectable using NMR or ESR spectroscopy. The labeled polypeptides and constructs of the present invention can be used, for example, in in vitro, in vivo or on-site assays (including immunoassays known per se, such as ELISA, RIA, EIA and "sandwich assays", etc.), as well as in vitro assays. In vivo diagnostic and imaging purposes will depend on the specific marker selection. As will be appreciated by those skilled in the art, another modification may include the introduction of a chelating group, for example one that chelates one of the metals or metal cations mentioned above. Suitable chelating groups include, for example, but are not limited to, diethylenetriaminepentaacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA). Yet another modification may include the introduction of a functional moiety that is part of a specific binding pair, such as a biotin-(streptavidin) binding pair. Such functional moieties can be used to link the polypeptide of the invention to another protein, polypeptide or chemical compound that binds to the other half of the binding pair, ie, by forming a binding pair. For example, a construct or polypeptide of the invention can be conjugated to biotin and linked to another protein, polypeptide, compound or carrier conjugated to avidin or streptavidin. For example, such conjugated constructs or polypeptides of the invention may be used as reporters, for example, in diagnostic systems, where the detectable signal-generating agent is conjugated to avidin or streptavidin. Such binding pairs may also be used, for example, to bind the constructs or polypeptides of the invention to carriers, including carriers suitable for pharmaceutical purposes. One non-limiting example is the liposome formulation described by Cao and Suresh (Journal of Drug Targeting 8: 257, 2000). Such binding pairs may also be used to link therapeutically active agents to the polypeptides of the invention. Other possible chemical and enzymatic modifications will be apparent to those skilled in the art. Such modifications may also be introduced for research purposes (eg, to study function-activity relationships). See, for example, Lundblad and Bradshaw (Biotechnol. Appl. Biochem. 26: 143-151, 1997). Preferably the constructs, polypeptides and/or derivatives are such that they have an affinity (with (real or apparent) K_D Value, (real or apparent) K_A value,k_on Association rate and/or k_off dissociation rate or alternatively in terms of IC₅₀ Values appropriately measured and/or expressed as further described herein) bind to MMP13. The constructs and/or polypeptides and derivatives thereof of the invention may also be in substantially isolated form (as defined herein). In one aspect, the invention relates to a construct of the invention comprising or consisting essentially of an ISVD according to the invention or a polypeptide according to the invention and which further comprises optionally linked via one or more peptide linkers one or more other groups, residues, moieties or binding units. In one aspect, the invention relates to constructs of the invention, wherein one or more other groups, residues, moieties or binding units are selected from the group consisting of: polyethylene glycol molecules, serum proteins or Its fragments, binding units that can bind to serum proteins, Fc portions and small proteins or peptides that can bind to serum proteins. Linkers In a construct of the invention (such as a polypeptide of the invention), two or more building blocks (such as ISVD) and optionally one or more other groups, drugs, agents, residues, moieties or binding units They may be linked to each other directly (as for example described in WO 99/23221) and/or may be linked to each other via one or more suitable spacers or linkers or any combination thereof. Spacers or linkers suitable for use in multivalent and multispecific polypeptides will be apparent to those skilled in the art, and may generally be any linker or spacer used in the art to link amino acid sequences. Preferably the linker or spacer is suitable for constructing constructs, proteins or polypeptides for medical use. For example, a polypeptide of the invention may be, for example, a trivalent, trispecific polypeptide comprising a building block that binds to MMP13 (such as an ISVD), a CAP building block (such as an ISVD that binds to aggrecan), and possibly another Building blocks (such as a third ISVD), wherein the first, second and third building blocks (such as an ISVD) are optionally linked via one or more and in particular via 2 linker sequences. The invention also provides constructs of the invention comprising a first ISVD that binds to MMP13 and a possible second ISVD and/or a possible third ISVD and/or a possible fourth ISVD that binds aggrecan. body or polypeptide, wherein the first ISVD and/or the second ISVD and/or the possible third ISVD and/or the possible fourth ISVD are linked via a linker, in particular via 3 linkers . Some particularly preferred linkers include those used in this technology to link antibody fragments or antibody domains. These include linkers mentioned in the general background cited above, as well as linkers used, for example, in the art to construct diabodies or ScFv fragments (however, in this regard, it should be noted that in bifunctional Linker sequences used in functional antibodies and in ScFv fragments should have permissive V_H and V_L The length, degree of flexibility and other properties of the domains that together form a complete antigen-binding site are not particularly limited as to the length or flexibility of the linkers used in the polypeptides of the invention, as each ISVD (such as Nanobodies ) itself forms a complete antigen-binding site). For example, the linker may be a suitable amino acid sequence, and particularly an amino acid sequence of between 1 and 50, preferably between 1 and 30, such as between 1 and 10 amino acid residues. . Some preferred examples of such amino acid sequences include, for example, the type described in WO 99/42077 (gly_x ser_y )_z gly-ser linkers (such as (gly₄ ser)₃ or(gly₃ ser₂ )₃ ) and the GS30, GS15, GS9 and GS7 linkers described in the Ablynx applications referred to herein (see e.g. WO 06/040153 and WO 06/122825), as well as hinge-like regions such as naturally occurring heavy chain antibodies or a hinge region of similar sequence (such as that described in WO 94/04678). Preferred linkers are described in Table C. Some other particularly good linkers are polyalanine (such as AAA), and the linkers GS30 (SEQ ID NO:85 in WO 06/122825) and GS9 (SEQ ID NO:84 in WO 06/122825) . Other suitable linkers generally include organic compounds or polymers, particularly those in proteins suitable for pharmaceutical use. For example, poly(ethylene glycol) moieties are used to link antibody domains, see eg WO 04/081026. It is within the scope of the present invention that the length, degree of flexibility, and/or other characteristics of the linker used (although not critical as they are often the linkers used for ScFv fragments) have a significant impact on the final construct of the invention ( Properties of polypeptides such as those of the invention may have some impact, including but not limited to affinity, specificity or retention for one or more of MMP13 or other antigens. Based on the disclosure herein, one skilled in the art will be free to determine, after some limited routine experimentation, the optimal linker for a particular construct of the invention (such as a polypeptide of the invention). For example, in multivalent polypeptides of the invention that include building blocks, ISVDs, or Nanobodies directed against MMP13 and other targets, the length and flexibility of the linker are preferably such that they allow for the presence of the linkers of the invention in the polypeptide. Each building block (such as an ISVD) binds to its cognate targets (eg, an antigenic determinant on each target). Based on the disclosure herein, one skilled in the art is free to determine, after some limited routine experimentation, the optimal linker for a particular construct of the invention (such as a polypeptide of the invention). It is also within the scope of the invention that the use of linkers confer one or more other advantageous properties or functionality to the constructs of the invention (such as the polypeptides of the invention) and/or provide for the formation of derivatives and/or use At one or more sites for attachment of functional groups (eg, as described herein for derivatives of ISVD of the invention). For example, linkers containing one or more charged amino acid residues can provide improved hydrophilic properties, while linkers that form or contain small epitopes or tags can be used for detection, identification and/or purification purposes. Based on the disclosure herein, one skilled in the art will be free to determine, after some limited routine experimentation, the optimal linker for a particular polypeptide of the invention. Finally, when two or more linkers are used in a construct of the invention (such as a polypeptide), the linkers may be the same or different. Based on the disclosure herein, one skilled in the art is free to determine, after some limited routine experimentation, the optimal linker for a particular construct and polypeptide of the invention. For ease of expression and production, constructs of the invention (such as polypeptides of the invention) are often linear polypeptides. However, the invention in its broadest sense is not limited thereto. For example, when a construct of the invention (such as a polypeptide of the invention) contains three or more building blocks, ISVDs or Nanobodies, it is possible that these may be linked using a linker with three or more "arms" , each "arm" is linked to a building block, ISVD or nanobody to provide a "star" construct. Although usually second best, it is also possible to use circular structures. Accordingly, the invention relates to constructs of the invention, such as polypeptides of the invention, wherein the ISVDs are linked to each other either directly or via a linker. The invention therefore relates to constructs of the invention, such as polypeptides of the invention, wherein the first ISVD and/or the second ISVD and/or a possible ISVD binding to serum albumin are linked via a linker. Therefore, the present invention relates to a construct of the invention, such as a polypeptide of the invention, wherein the linker is selected from the group consisting of: A3, 5GS, 7GS, 8GS, 9GS, 10GS, 15GS, 18GS, 20GS, 25GS, 30GS, 35GS, 40GS, G1 hinge, 9GS-G1 hinge, vicuña long hinge, and G3 hinge, such as those presented in Table C. Therefore, the present invention relates to a construct of the invention, such as a polypeptide of the invention, wherein the polypeptide is selected from the group shown in Table A-3 and Table F, for example, selected from the group consisting of: SEQ ID NO :164 to 165, 160, 161, 162, 163, and SEQ ID NO: 176, 192 and 175 to 191 (i.e. 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190 or 191). Preparation The present invention further relates to methods of making the constructs, polypeptides, ISVDs, nucleic acids, host cells and compositions described herein. Multivalent polypeptides of the invention can generally be prepared by a method comprising at least the following steps: suitably linking an ISVD and/or a monovalent polypeptide of the invention to one or more additional ISVDs, optionally via one or more suitable linkers. , in order to provide the multivalent polypeptide of the present invention. The polypeptides of the invention can also be prepared by a method generally comprising at least the following steps: providing a nucleic acid encoding a polypeptide of the invention, expressing the nucleic acid in a suitable manner, and recovering the expressed polypeptide of the invention. These methods may be performed in known manner per se apparent to those skilled in the art, for example based on the methods and techniques further described herein. The method for preparing the multivalent polypeptide of the present invention may comprise at least the following steps: linking two or more ISVDs of the present invention together with, for example, one or more linkers in a suitable manner. The ISVDs (and linkers) of the present invention may be coupled by any method known in the art and as further described herein. Preferred techniques involve linking nucleic acid sequences encoding ISVDs of the invention (with linkers) to prepare genetic constructs expressing multivalent polypeptides. Techniques for linking amino acids or nucleic acids will be apparent to those skilled in the art, and reference is made to standard manuals such as Sambrook et al. and Ausubel et al., mentioned above, and to the Examples below. Therefore, the present invention also relates to the use of the ISVD of the invention to prepare multivalent polypeptides of the invention. Methods of preparing multivalent polypeptides comprise linking an ISVD of the invention to at least one additional ISVD of the invention, optionally via one or more linkers. The ISVDs of the invention are then used as binding domains or building blocks to provide and/or prepare compounds containing 2 (eg as a bivalent polypeptide), 3 (eg as a trivalent polypeptide), 4 (eg as a tetravalent polypeptide) or more A multivalent polypeptide of multiple (e.g., in the form of a multivalent polypeptide) building blocks. In this regard, ISVDs of the invention can be used as binding domains or binding units to provide and/or prepare multivalent (such as bivalent, trivalent or tetravalent) compounds of the invention comprising 2, 3, 4 or more building blocks. Peptides. Accordingly, the present invention also relates to the use of an ISVD polypeptide of the invention (as described herein) to prepare a multivalent polypeptide. Methods for preparing multivalent polypeptides comprise linking an ISVD of the invention to at least one additional ISVD of the invention, optionally via one or more linkers. The polypeptides and nucleic acids of the invention may be prepared in a manner known per se, as will be apparent to those skilled in the art from the further description herein. For example, the polypeptides of the invention may be prepared in any manner known per se for the preparation of antibodies and in particular for the preparation of antibody fragments, including but not limited to (single) domain antibodies and ScFv fragments. Some preferred, but non-limiting, methods for preparing polypeptides and nucleic acids include the methods and techniques described herein. Methods for producing a polypeptide of the invention may comprise the steps of: - Expression in a suitable host cell or host organism (also referred to herein as a "host of the invention") or in a nucleic acid encoding the polypeptide of the invention ( In another suitable expression system, also referred to herein as "nucleic acid of the invention"), optionally follows: - isolation and/or purification of the polypeptide of the invention thus obtained. In particular, this method may comprise the following steps: - culturing and/maintaining a host of the invention under conditions such that the host of the invention expresses and/or produces at least one polypeptide of the invention; optionally followed by: - isolating and/or Or purify the polypeptide of the invention thus obtained. Accordingly, the invention also relates to nucleic acids or nucleotide sequences encoding polypeptides, ISVDs or constructs of the invention (also referred to as "nucleic acids of the invention"). The nucleic acids of the invention may be in the form of single- or double-stranded DNA or RNA. According to one embodiment of the invention, the nucleic acids of the invention are in substantially isolated form, as defined herein. The nucleic acids of the invention are also in the form of, present in and/or part of a vector, eg an expression vector such as a plasmid, cosmid or YAC, which may also be in a substantially isolated form. Therefore, the present invention also relates to expression vectors comprising the nucleic acid or nucleotide sequence of the invention. The nucleic acids of the invention can be prepared or obtained in a manner known per se based on the information given herein about the polypeptides of the invention and/or can be isolated from suitable natural sources. As will also be apparent to those skilled in the art, a number of nucleotide sequences (such as at least two nucleic acids encoding an ISVD of the invention and, for example, a nucleic acid encoding one or more linkers) may also be linked together in a suitable manner to prepare the invention. Invention of nucleic acids. Techniques for producing nucleic acids of the invention will be apparent to those skilled in the art and may include, for example, but are not limited to, automated DNA synthesis, site-guided mutagenesis, combination of two or more naturally occurring and/or synthetic sequences (or two or more parts), introduction of mutations resulting in the expression of a truncated expression product, introduction of one or more restriction sites (e.g., the creation of gene cassettes and/or regions that can be readily digested and/or ligated using suitable restriction enzymes), and/or introduce mutations by means of PCR reactions using one or more "mismatch" primers. These and other techniques will be apparent to those skilled in the art and reference is made to standard manuals such as Sambrook et al. and Ausubel et al., mentioned above, and to the examples below. In a preferred but non-limiting embodiment, the genetic construct of the invention comprises a) at least one nucleic acid of the invention; b) together with one or more regulatory elements (such as a promoter and optionally a suitable terminator). operably linked; and optionally also c) one or more other elements of the genetic construct known per se; wherein the terms "regulatory element", "promoter", "terminator" and "operably linked" have their customary meaning in this technology. Genetic constructs of the invention may generally be provided by suitably linking the nucleotide sequence of the invention to one or more of the other elements described above, for example using techniques described in general manuals, such as those mentioned above Sambrook et al. and Ausubel et al. The nucleic acids of the invention and/or the genetic constructs of the invention can be used to transform host cells or host organisms, that is, to express and/or produce the polypeptides of the invention. Suitable hosts or host cells will be apparent to those skilled in the art and may, for example, be any suitable fungal, prokaryotic or eukaryotic cell or cell strain or any suitable fungal, prokaryotic or (non-human) eukaryotic organism; and All other host cells or (non-human) hosts known per se for the expression and production of antibodies and antibody fragments, including but not limited to (single) domain antibodies and ScFv fragments, will be apparent to those skilled in the art. Reference is also made to the general background cited above, and to eg WO 94/29457; WO 96/34103; WO 99/42077; Frenken et al. (Res Immunol. 149: 589-99, 1998); Riechmann and Muyldermans (1999) , mentioned above; van der Linden (J. Biotechnol. 80: 261-70, 2000); Joosten et al. (Microb. Cell Fact. 2: 1, 2003); Joosten et al. (Appl. Microbiol. Biotechnol. 66 : 384-92, 2005); and further references cited in this article. In addition, the polypeptides of the invention can also be expressed and/or produced in cell-free expression systems, and suitable examples of such systems will be apparent to those skilled in the art. Suitable techniques for transforming the host or host cells of the invention will be apparent to those skilled in the art and may depend on the intended host cell/host organism and the genetic construct intended to be used. Refer again to the manuals and patent applications mentioned above. Transformed host cells (which may be in the form of stable cell lines) or host organisms (which may be in the form of stable mutants or strains) constitute further aspects of the invention. The invention therefore relates to a host or host cell comprising a nucleic acid according to the invention or an expression vector according to the invention. Preferably such host cells or host organisms are such that they express or (at least) are capable of expressing (e.g. under suitable conditions) a polypeptide of the invention (and in the case of a host organism: in at least one cell, part thereof , tissues or organs). The invention also includes other generations, progeny and/or progeny of the host cell or host organism of the invention, which may be obtained, for example, by cell division or by sexual or asexual reproduction. In order to produce/obtain the expression of a polypeptide of the invention, the transformed host cell or the transformed host organism can generally be maintained, maintained under conditions and/or in which the (desired) polypeptide of the invention can be expressed/produced. cultured under conditions. Suitable conditions will be apparent to those skilled in the art and will generally depend on the host cell/host organism used, as well as the regulatory elements controlling the expression of the (relevant) nucleotide sequence of the invention. Reference is again made to the manuals and patent applications mentioned in the paragraph above regarding the genetic constructs of the present invention. The polypeptide of the invention can then be isolated from the host cell/host organism and/or from the culture medium in which the host cell or host organism is cultured, using protein isolation and/or purification techniques known per se, such as (preparative) Chromatography and/or electrophoresis techniques, differential precipitation techniques, affinity techniques (for example, the use of specific, cleavable amino acid sequences fused to the polypeptide of the invention) and/or preparative immunology techniques (that is, the use of specific and cleavable amino acid sequences fused to the polypeptide of the invention) Antibodies to isolated polypeptides). In one aspect, the invention relates to a method of producing a construct, polypeptide or ISVD according to the invention, comprising at least the steps of: (a) expressing in a suitable host cell or host organism or in another suitable expression system A nucleic acid sequence according to the invention; optionally followed by (b) isolating and/or purifying a construct, polypeptide or ISVD according to the invention. In one aspect, the invention relates to compositions comprising a construct, polypeptide, ISVD or nucleic acid according to the invention. Agents (Uses of ISVDs, Polypeptides, and Constructs of the Invention) As mentioned herein, there is still a need for safe and effective OA agents. Based on unconventional screening, characterization and combination strategies, the inventors identified ISVDs that bind and inhibit MMP13. These MMP13 binding agents performed exceptionally well in in vitro and in vivo experiments. Furthermore, the ISVD of the present invention also proved to be significantly more effective than the comparative molecules. The present invention therefore provides ISVDs and polypeptides that antagonize MMPs, particularly MMP13, with improved prophylactic, therapeutic and/or pharmacological properties compared to comparative molecules, including a safer profile. Additionally, when these MMP13 binding agents are linked to the CAP building blocks, they have increased retention in joints and, on the other hand, retained activity. In one aspect, the invention relates to a composition according to the invention, an ISVD according to the invention, a polypeptide according to the invention and/or a construct according to the invention for use as a medicament. In another aspect, the present invention relates to the use of the ISVD, polypeptide and/or construct of the present invention to prepare a pharmaceutical composition for preventing and/or treating diseases related to at least MMP13; and/or For use in one or more of the treatments mentioned herein. The present invention also relates to the use of the ISVD, polypeptide, compound and/or construct of the present invention to prepare pharmaceutical compositions for prevention and/or treatment by modulating the activity of MMP (preferably MMP13), for example At least one disease or disease that is prevented and/or treated by inhibiting the degradation of aggrecan and/or collagen. The present invention also relates to the use of the ISVD, polypeptide, compound and/or construct of the present invention to prepare a pharmaceutical composition. The pharmaceutical composition is used for prevention and/or treatment by administering the ISVD, polypeptide, compound and/or construct of the present invention to a patient. At least one disease, disorder or symptom for the prevention and/or treatment of compounds and/or constructs. The invention further relates to the ISVDs, polypeptides, compounds and/or constructs of the invention or pharmaceutical compositions comprising them for the prevention and/or treatment of at least one MMP13-related disease. The MMP13 binding agents of the present invention are expected to be useful in various diseases affecting cartilage, such as joint pathologies and chondrodystrophies, arthritic diseases such as osteoarthritis, rheumatoid arthritis, gouty arthritis, psoriatic arthritis, traumatic arthritis, Rupture or detachment, achondroplasia, costochondritis, aplasia of the spine, disc herniation, lumbar disc degenerative disease, degenerative joint disease, and relapsing polychondritis, osteochondritis dissecans, and aggrecan pathology (This article is often expressed as "MMP13-related diseases"). In one aspect, the invention relates to compositions, ISVDs, polypeptides and/or constructs according to the invention for use in the treatment or prevention of symptoms of MMP13-related diseases, such as for use in joint pathologies and chondrodystrophies, arthritis Diseases such as osteoarthritis, rheumatoid arthritis, gouty arthritis, psoriatic arthritis, traumatic rupture or detachment, achondroplasia, costochondritis, spondylosis, disc herniation, lumbar degenerative disc disease , degenerative joint diseases and relapsing polychondritis dissecans, and aggrecan lesions. More preferably the disease or disorder is arthritic disease, and most preferably osteoarthritis. In one aspect, the invention relates to use in the prevention or treatment of joint pathology and chondrodystrophies, arthritic diseases such as osteoarthritis, rheumatoid arthritis, gouty arthritis, psoriatic arthritis, traumatic rupture or Methods for detachment, achondroplasia, costochondritis, aplasia of the vertebrae, disc herniation, degenerative lumbar disc disease, degenerative joint disease and relapsing polychondritis, wherein the method includes treatment for individuals in need (for those with A person in need thereof) is administered a pharmaceutically active amount of at least a composition, immunoglobulin, polypeptide or construct according to the invention. More preferably the disease is an arthritic disease, and most preferably osteoarthritis. In one aspect, the invention relates to the use of a pharmaceutical composition prepared according to the ISVD, polypeptide, composition or construct of the invention for the treatment or prevention of diseases or disorders such as joint pathology and cartilage nutrition. Unhealthy, arthritic diseases such as osteoarthritis, rheumatoid arthritis, gouty arthritis, psoriatic arthritis, traumatic rupture or detachment, achondroplasia, costochondritis, aplasia of the vertebrae, disc herniation, lumbar Degenerative disc disease, degenerative joint disease, and relapsing polychondritis, osteochondritis dissecans, and aggrecan lesions. More preferably the disease or disorder is arthritic disease, and most preferably osteoarthritis. The constructs and/or polypeptides of the present invention can reduce or inhibit serine protease family members, autolyses, matrix metalloproteinases (MMPs)/matrix metalloproteinases that degrade aggrecan by binding to aggrecan. (Matrixin) or disintegrin (Disintegrin) and metalloproteinase (ADAMTS) with platelet thrombin (Thrombospondin), MMP20, ADAMTS5 (Aggrecanase)-2), ADAMTS4 (aggrecanase-1 ) and/or the activity of ADAMTS11. In the context of the present invention, the term "prevention and/or treatment" includes not only the prevention and/or treatment of a disease, but also generally the prevention of the onset of a disease, the slowing down or reversal of the progression of a disease, the prevention or slowing down of one of the or The onset of multiple symptoms, reduction and/or alleviation of one or more symptoms associated with the disease, reduction in the severity and/or duration of the disease and/or any symptoms associated with the disease and/or prevention of further increase in the disease and/or association with the disease The severity of any symptoms associated with the disease, the prevention, reduction or reversal of any physiological damage caused by the disease, and generally any pharmacological effects that are beneficial to the patient being treated. Dosage regimens are determined by the attending physician and clinical factors. As is well known to those skilled in the art of medicine, the dosage for any given patient will depend on many factors, including the patient's size, weight, body surface area, age, the specific compound being administered, and the type of polypeptide (including antibodies) being used. Activity, time and route of administration, general health, and combination with other treatments or treatments. The protein pharmaceutical active may be present in an amount between 1 gram and 100 mg/kg body weight per dose; however, doses below or above this exemplary range are also contemplated. If the regimen is a continuous infusion, it may range from 1 picogram to 100 mg per kilogram of body weight per minute. ISVDs, polypeptides or constructs of the invention may be used at a concentration of, for example, 0.01, 0.1, 0.5, 1, 2, 5, 10, 20 or 50 pg/ml to inhibit and/or neutralize by at least about 50%, preferably is 75%, more preferably 90%, 95% or at most 99%, and most preferably about 100% (substantially complete) of the biological function of MMP13, as assayed by methods well known in the art. ISVDs, polypeptides or constructs of the invention may be used, for example, at a concentration of 1, 2, 5, 10, 20, 25, 30, 40, 50, 75, 100, 200, 250 or 500 ng/mg of cartilage to inhibit and /or neutralize at least about 50%, preferably 75%, more preferably 90%, 95% or at most 99%, and most preferably about 100% (substantially completely) of the biological function of MMP13, as in this case Tested by methods well known in the art. Typically, treatment regimens include the administration of a pharmaceutically effective amount or dose of one or more ISVDs, polypeptides and/or constructs of the invention or comprise one or more compositions thereof. The specific amount or dose to be administered can be determined by the clinician based upon the factors cited above. Useful dosages of the constructs, polypeptides and/or ISVDs of the invention can be determined by comparing their in vitro and in vivo activities in animal models. Methods for extrapolating effective doses in mice and other animals to humans are known in the art; eg US 4,938,949. Typically the clinician can depend on the specific disease, disorder or condition to be treated, the efficacy of the specific ISVD, polypeptide and/or construct of the invention to be used, the route of administration and the specific pharmaceutical formulation or composition to be used. to determine the appropriate daily dose. The amount of constructs, polypeptides and/or ISVDs of the invention required for treatment will vary not only with the particular immunoglobulin, polypeptide, compound and/or construct selected, but also with the route of administration, type of treatment. It will vary depending on the nature of the symptoms and the age and condition of the patient, and is ultimately at the discretion of the attending physician or clinician. The dosage of the constructs, polypeptides and/or ISVDs of the invention will also vary depending on the target cell, tissue, graft, joint or organ. The desired dose may be presented as a single dose or as divided doses administered conveniently at appropriate intervals, for example, as two, three, four or more doses per day. The sub-doses themselves may be further divided, for example, into a number of individual, not strictly spaced administrations. Preferably the dosage is administered once a week or even less frequently, such as once every two weeks, once every three weeks, once a month or even once every two months. Dosage regimens may include long-term treatment. "Long-term" means a duration of at least two weeks and preferably weeks, months or years. Necessary modifications within this dosage range can be determined by one of ordinary skill in the art using only routine experimentation as given by the guidance herein. See Remington’s Pharmaceutical Sciences (Martin, E.W., ed.4), Mack Publishing Co., Easton, PA. The dosage may also be adjusted by the individual physician in case of any complications. Typically the ISVDs, polypeptides and/or constructs of the invention are used in the above methods. However, it is within the scope of the invention to use combinations of two or more ISVDs, polypeptides and/or constructs of the invention. The ISVDs, polypeptides and/or constructs of the present invention can be used in combination with one or more additional pharmaceutically active compounds or main drugs that may or may not lead to a synergistic effect, ie, as a combination treatment regimen. Pharmaceutical compositions may also contain at least one additional active agent, such as one or more additional antibodies or antigen-binding fragments thereof, peptides, proteins, nucleic acids, organic and inorganic molecules. Furthermore, clinicians can select such additional compounds or main drugs, as well as appropriate combination treatment options, based on the factors cited above and their professional judgment. In particular, the ISVDs, polypeptides and/or constructs of the present invention can be used in combination with other pharmaceutically active compounds or main drugs that are or can be used to prevent and/or treat the diseases, disorders and symptoms cited above, and the results may or Synergy effects may not be obtained. Examples of such compounds and agents, as well as routes, methods, and pharmaceutical formulations or compositions for administering the same, are known to clinicians. When two or more substances or agents are used as part of a combination treatment regimen, they may be administered via the same route of administration or via different routes of administration, at substantially the same time or at different times (e.g. Administered substantially simultaneously, sequentially, or on an alternating schedule). When the substances or active ingredients are administered simultaneously by the same route of administration, they may be administered as different pharmaceutical formulations or compositions or as part of a combined pharmaceutical formulation or composition, to those skilled in the art All is clear. Furthermore, when two or more substances or agents are used as part of a combination treatment regimen, each substance or agent may be used in the same amount and on the same basis as when the compound or agent is used alone. regimen, and this combination may or may not result in synergistic effects. However, when the combined use of two or more active substances or active substances results in a synergistic effect, it is also possible to reduce the amount of one, more or all of the substances or active ingredients to be administered, while still achieving the desired effect. Therapeutic effect. This may, for example, serve to avoid, limit or reduce undesirable side effects associated with one or more of the substances or principal drugs when used in their customary amounts, while still obtaining the desired medicinal or therapeutic effect. The effectiveness of a treatment regimen used in accordance with the present invention may be determined and/or tracked in any manner known per se for the disease, disorder or condition involved, as will be apparent to the clinician. Clinicians are able, when appropriate and on a case-by-case basis, to alter or modify a particular treatment plan in order to achieve the desired therapeutic effect, avoid, limit or reduce undesirable side effects, and/or achieve the desired therapeutic effect on the one hand and avoid, strike the right balance between limiting or reducing undue side effects. Treatment regimens are typically followed until the desired therapeutic effect is achieved and/or as long as the desired therapeutic effect is maintained. Furthermore, this can be determined by the clinician. Therefore, in another aspect, the invention relates to a pharmaceutical composition comprising at least one construct of the invention, at least one polypeptide of the invention, at least one ISVD of the invention or at least one nucleic acid of the invention and at least one suitable carriers, diluents or excipients (i.e. suitable for pharmaceutical use) and optionally one or more additional active substances. In a particular aspect, the invention relates to pharmaceutical compositions comprising a construct, polypeptide, ISVD or nucleic acid according to the invention (preferably at least one of the following: SEQ ID NO: 111, 11, 112, 12 ,109,9,110,10,1,13,14,15,16,17,18,19,20,21,22,2,3,4,5,6,7,8,160-165 (also That is, SEQ ID NO: 160, 161, 162, 163, 164 or 165) and 176 to 192 (that is, SEQ ID NO: 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 or 192)) and at least one suitable carrier, diluent or excipient (ie suitable for pharmaceutical use) and optionally one or more additional active substances. The individual to be treated can be any warm-blooded animal, but particularly mammals, and more particularly humans. In veterinary applications, an individual to be treated includes any animal kept for commercial purposes or kept as a pet. As will be appreciated by those skilled in the art, individuals to be treated are particularly those suffering from or at risk of the diseases, disorders and conditions mentioned herein. Therefore, in a preferred embodiment of the present invention, a pharmaceutical composition comprising the polypeptide of the present invention is used in medicine or diagnostics. Preferably the pharmaceutical compositions are for use in human medicine, but they may also be used for veterinary purposes. Furthermore, in these pharmaceutical compositions, one or more immunoglobulins, polypeptides, compounds and/or constructs of the present invention, or nucleotides encoding them and/or pharmaceutical compositions containing them can also be used. Suitably combined with one or more other active ingredients (such as those mentioned herein). The invention also relates to use in vitro (e.g. in vitro or in cellular assays) or in vivo (e.g. in unicellular or multicellular organisms, and in particular in mammals, and more particularly in humans, such as in compositions (such as, but not limited to, pharmaceutical compositions or formulations, as further described herein) in humans who are at risk for or suffering from the diseases, disorders, or symptoms of the invention). It should be understood that treatment referred to includes both treatment of established symptoms and preventive treatment, unless otherwise expressly stated. The constructs, polypeptides and/or ISVDs of the invention generally used for pharmaceutical purposes may be formulated into pharmaceutical preparations or compositions, which comprise at least one construct, polypeptide and/or ISVD of the invention and at least one pharmaceutically acceptable carrier. agents, diluents or excipients and/or adjuvants, and optionally one or more pharmaceutically active polypeptides and/or compounds. By way of non-limiting example, such formulations may be in a form suitable for oral administration, parenteral administration (such as intravenous, intramuscular or subcutaneous injection or intravenous infusion), local administration (such as intra-articular administration). administration), inhalation administration, transdermal patch, implant, suppository, etc., among which intra-articular administration is preferred. Such suitable administration forms will be apparent to those skilled in the art and may be solid, semi-solid or liquid depending on the mode of administration as well as the methods and vehicles used to prepare them, and are further described herein. Such pharmaceutical preparations or compositions are generally referred to herein as "pharmaceutical compositions." Disintegrating agents, binding agents, fillers and lubricants may be mentioned as exemplary excipients. Examples of disintegrants include agar, algin, calcium carbonate, cellulose, colloidal silica, gum, magnesium aluminum silicate, methylcellulose and starch. Examples of binding agents include microcrystalline cellulose, hydroxymethylcellulose, hydroxypropylcellulose and polyvinylpyrrolidone. Examples of fillers include calcium carbonate, calcium phosphate, calcium sulfate, calcium carboxymethyl cellulose, cellulose, dextrin, dextrose, fructose, lactitol, lactose, magnesium carbonate, magnesium oxide, maltitol, maltodextrin , maltose, sorbitol, starch, sucrose, sugar and xylitol. Examples of lubricants include agar, ethyl oleate, ethyl laurate, glycerin, glyceryl palmitostearate, hydrogenated vegetable oil, magnesium oxide, stearates, mannitol, poloxamer, dihydrogen alcohol, sodium benzoate, sodium lauryl sulfate, sodium stearate, sorbitol and talc. Useful stabilizers, preservatives, wetting and emulsifying agents, consistency improvers, odor improvers, osmotic pressure modifying salts, buffer substances, solubilizers, diluents, emollients, colorants and opacifying agents and antioxidants Considered as a pharmaceutical adjuvant. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting point Waxes, cocoa butter, water, alcohols, polyols, glycerin, vegetable oils and the like. Generally, the constructs, polypeptides and/or ISVDs of the invention may be formulated and administered in any suitable manner known per se. Reference is made to the general background art cited above (and in particular to WO 04/041862, WO 04/041863, WO 04/041865, WO 04/041867 and WO 08/020079) and to standard handbooks such as Remington’s Pharmaceutical Sciences, 18^th Ed., Mack Publishing Company, USA (1990), Remington, the Science and Practice of Pharmacy, 21st Edition, Lippincott Williams and Wilkins (2005); or the Handbook of Therapeutic Antibodies (S. Dubel, Ed.), Wiley, Weinheim , 2007 (see e.g. pp. 252 to 255). In a particular aspect, the invention relates to pharmaceutical compositions comprising a construct, polypeptide, ISVD or nucleic acid according to the invention and further comprising at least one pharmaceutically acceptable carrier, diluent or excipient and/or Adjuvants, and optionally include one or more additional pharmaceutically active polypeptides and/or compounds. The constructs, polypeptides and/or ISVDs of the invention may be formulated and administered in any manner known per se for conventional antibodies and antibody fragments (including ScFv and diabodies) and other pharmaceutically active proteins. Such formulations and methods for preparing them will be apparent to those skilled in the art, and include, for example, those suitable for parenteral administration (e.g., intravenous, intraperitoneal, subcutaneous, intramuscular, intravascular, intraarterial , intraspinal, intranasal or intrabronchial administration), but is also a preferred formulation suitable for local (such as intra-articular, transdermal or intradermal) administration. Formulations for topical or parenteral administration may, for example, be sterile solutions, suspensions, dispersions or emulsions suitable for infusion or injection. Suitable carriers or diluents for such formulations include, for example, those mentioned on page 143 of WO 08/020079. Usually aqueous solution or suspension is preferred. The constructs, polypeptides and/or ISVDs of the invention may also be administered using delivery methods known from gene therapy, see for example U.S. Patent No. 5,399,346, which is incorporated by reference for its gene therapy delivery methods. Using gene therapy delivery methods, primary cells transfected with genes encoding constructs, polypeptides, and/or ISVDs of the invention can be additionally transfected with tissue-specific promoters to target specific organs, tissues, grafts, joints, or cells, and may additionally be transfected with signals and stabilizing sequences for subcellular localization expression. According to additional aspects of the invention, the polypeptides of the invention can be used for additional applications in vivo and in vitro. For example, the polypeptides of the invention may be used for diagnostic purposes, such as in assays designed to detect and/or quantify the presence of MMP13 and/or purify MMP13. Peptides can also be tested in animal models of specific diseases and used to conduct toxicity, safety and dosage studies. Finally, the invention relates to a kit comprising at least one polypeptide according to the invention, at least one nucleic acid sequence encoding said component, vector or vector system according to the invention and/or a host cell according to the invention. It is contemplated that the kit may be supplied in different forms, for example as a diagnostic kit. The present invention is now further explained with the help of the following non-limiting preferred aspects, examples and figures. The entire contents of all references cited throughout this application (including literature references, issued patents, published patent applications, and patent applications also pending) are hereby expressly incorporated by reference. This article is specifically for the guidance mentioned above. The sequence is disclosed in the main body of the description and in the individual sequence listing according to WIPO Standard ST.25. The SEQ ID specified by a specific number should be the same in the main body of the description and in the individual sequence listing. To illustrate by example, SEQ ID no.:1 should define the same sequence in both the main body of the description and the individual sequence listing. If there is an inconsistency between the sequence definition in the main body of the specification and the individual sequence listing (if, for example, SEQ ID no.:1 in the main body of the description incorrectly corresponds to SEQ ID no.:2 in the individual sequence listing), it should be understood that in the application The specific sequences mentioned in the application (especially in specific embodiments) are the sequences mentioned in the main body of the application and not in the individual sequence listings. In other words, inconsistencies between sequence definitions/names in the main body of the specification and the individual sequence listings should be disclosed by correcting the individual sequence listings into the main body of the application including the inventive content, embodiments, drawings and patentable scope. sequence and their names. EXAMPLES The following examples illustrate the methods and products of the present invention. Appropriate modifications and adaptations of the described conditions and parameters commonly found in the molecular and cell biology arts will be apparent to those skilled in the art and are within the spirit and scope of the invention. 6.1 Methods 6.1.1 Llama immunization The Llama immunization method was performed according to standard procedures with varying amounts of antigen, type of adjuvant, and injection method used. These variations are detailed in the following paragraphs. All immunological experiments were approved by the local ethical committee. 6.1.2 Library construction cDNA was prepared using total RNA extracted from blood samples of all llamas/alpacas immunized with MMP13. The nucleotide sequence encoding the nanobody was amplified from cDNA in a one-step RT-PCR reaction and ligated into the corresponding restriction site of the phagemid vector pAX212, and the ligated product was then used to transform E. coli strain TG via electroporation. -1. The NNK library was generated by overlap extension PCR with degenerate primers. The PCR product was cloned in an expression vector (pAX129) and transformed into E. coli TG-1 competent cells. In frame with the Nanobody coding sequence, the vector encodes C-terminal FLAG₃ - and His₆ -Tag. The selected strains of concern have been sequence verified. 6.1.3 Selection Phage display libraries were probed using recombinant MMP13. Different rounds of selection use different antigens, as detailed in the Results section (i.e. Example 6.2) and subsequently. Selection consists of incubating the antigen with library phage particles for 2 hours in PBS supplemented with 2% Marvel and 0.05% Tween 20. Biotinylated antigens were captured using streptavidin-coated magnetic beads (Invitrogen, 112-05D). Non-biotinylated antigen was plated on MaxiSorp plates (Nunc, 430341). Unbound phage were washed out (PBS supplemented with 0.05% Tween 20); bound phage system was eluted by adding trypsin (1 mg/ml in PBS) for 15 minutes. Exponentially growing E. coli TG-1 cells were infected with lysed phage for phage rescue. Phage systems prepared from the output of selection are used as input for subsequent selection rounds. 6.1.4 Antigens directly coated by ELISA MaxiSorp plates (Nunc, 430341) were coated with human proMMP 13 overnight at 4°C, followed by blocking (PBS, 1% casein) for 1 hour at RT. After the wash step in PBS + 0.05% Tween 20, a 10-fold dilution of the periplasmic extract in PBS, 0.1% casein, 0.05% Tween 20 was added over 1 h at RT. Bound Nanobodies were detected with mouse anti-FLAG-HRP (Sigma (A8592)). Captured Antigen - Activated Human MMP13 ELISA - Activated Rat MMP13 ELISA - Activated Dog MMP13 ELISA MaxiSorp plates (Nunc, 430341) were coated with human MMP13 antibody mAb511 overnight at 4°C, followed by 1 hour blocking (PBS, 1% casein). After the wash step in PBS + 0.05% Tween20, 20 nM activated human, rat or dog MMP13 was added over 1 hour at RT. After the second wash step, 10-fold dilutions of periplasmic extracts or serial dilutions of purified nanobodies in PBS, 0.1% casein, 0.05% Tween 20 were added over 1 h at RT. Bound Nanobodies were detected with mouse anti-FLAG-HRP (Sigma (A8592)). 6.1.5 Fluorescent peptide assay The settings for the human, macaque, rat, dog and bovine MMP13 fluorescent peptide assay and the human MMP1 and MMP14 fluorescent peptide assay are briefly as follows: Combine activated MMP and fluorescent peptide substrates Mca-PLGL-Dpa-AR-NH2 (R&D Systems #ES001) and 1/5 dilutions of periplasmic extract or serial dilutions of purified nanobodies/positive controls (total volume = 20 μl, in assay buffer 50 mM Tris pH 7.5, 100 mM NaCl, 10 mM CaCl2, 0.01% Tween20) and incubated at 37°C for 2 hours. Use the linear increase in fluorescence (v0 - with incubation between 15 and 45 minutes) as a measure of enzymatic activity and the formula 100-100 (v0 in the presence of test Nanobodies/negative control Nanobodies (Cablys v0) in the presence of ) calculates % inhibition. 6.1.6 Collagen solubilization assay The setup of this assay is as follows: 250ng/ml immune-grade human collagen II (Chondrex #20052) and 5 nM activated MMP13 were dissolved in 100μl assay buffer (50 mM Tris-Cl pH 7.5, 100 mM NaCl, 10 mM CaCl2, 0.01% Tween-20). After incubation at 35°C for 1.5 hours, the reaction was neutralized with EDTA (10 μl of 30 mM stock). MMP13-cleaved collagen was further degraded with elastase at 38°C for 20 minutes to avoid degradation of collagen II (1/3 dilution of 10 μl provided in Type II Collagen Assay Kit (Chondrex #6009) Stock solution) re-annealing. The remaining intact collagen is tested by ELISA using the reagents provided in the Type II Collagen Assay Kit (Chondrex #6009). 6.1.7 Fluorescent Collagen Assay The setup of this assay is as follows: 100 μg/ml of type I DQ™ collagen from bovine skin (fluorescein conjugate; Molecular Probes #D-12060, lot number 1149062 ) with 10 nM activated MMP13 and serial dilutions of purified Nanobodies/positive controls in 40 μl assay buffer (50 mM Tris-Cl pH 7.5, 100 mM NaCl, 10 mM CaCl) at 37°C.₂ , 0.01% Tween-20) for 2 hours. Use the linear increase in fluorescence (v0 - with incubation between 15 and 45 minutes) as a measure of enzymatic activity and the formula 100-100 (v0 in the presence of test Nanobodies/negative control Nanobodies (Cablys v0) in the presence of ) calculates % inhibition. 6.1.8 TIMP-2 Competition Assay 50 μl of TIMP-2 (0.63 nM; R&D Systems #971-TM) was captured (1 hour at RT) coated with anti-human TIMP-2 antibody (R&D Systems #MAB9711). plate (2 μg/ml in PBS; overnight). During this capture, serial dilutions of 1.26 nM activated MMP-13-biotin with Nanobody/TIMP-3/MSC2392891A were mixed in 70 μl of assay buffer (50 mM Tris-Cl pH 7.5, 100 mM NaCl, 10 mM CaCl₂ , 0.01% Tween-20) pre-cultured. 50 μl of this mixture was added to the captured TIMP-2 and incubated for 1 hour at room temperature. MMP13-biotin was detected with 50 μl of Streptavidin-HRP (1:5000 DakoCytomation #P0397). 6.1.9 Thermal transfer assay (TSA) TSA was performed basically according to Ericsson et al. (2006 Anals of Biochemistry, 357: 289-298) with 5 μl of purified monovalent nanobodies. 6.1.10 Analytical size exclusion chromatography (analytical SEC) Analytical SEC experiments were performed on an Ultimate 3000 machine (Dionex) combined with a Biosep-SEC-3 (Agilent) column. 6.1.11 Forced oxidation: Nanobody samples (1mg/ml) were subjected to 10 mM H in PBS for 4 hours at RT and in the dark.₂ O₂ , while without H₂ O₂ Control samples were performed, and the buffer was subsequently converted to PBS using a Zeba desalted spin column (0.5 ml) (Thermo Scientific). The stressed and control samples were then analyzed by RPC at 70°C on a Zorbax 300SB-C3 column (Agilent Technologies) on a 1200 or 1290 series machine (Agilent Technologies). Oxidation of Nanobodies was quantified by determining the % peak area of pre-peaks that appear due to oxidative stress compared to the main protein peak. 6.1.12 Temperature stress Nanobody samples (1 to 2 mg/ml) were stored in PBS at -20°C (negative control), 25°C, and 40°C for 4 weeks. After this incubation period, the Nanobodies are digested with trypsin or LysC. The sample was then analyzed by RPC at 60°C on an Acquity UPLC BEH300-C18 column (Agilent Technologies) coupled to a Q-TOF mass spectrometer (6530 Accurate Mass Q-TOF (Agilent)) on a 1290 series machine (Agilent Technologies). and the peptide of the control sample. 6.2 Immunization method MMP13 is secreted in the inactive pro-form (proMMP13). This form is activated when the prodomain is cleaved, leaving the active enzyme consisting of the catalytic domain forming the catalytic pocket and the thrombin-like domain (PDB: 1PEX), which has been shown to act as a substrate for collagen II (Col II) The docking/interaction domain functions. It is speculated that the best area to inhibit the enzymatic activity of MMP13 may be the catalytic pocket. However, increasing the immune response against the catalytic bag has various important problems. First, in proMMP13, the prodomain obscures the catalytic pocket, so the catalytic pocket is not available to enhance immune responses. Second, activated MMP13 has a short half-life, mainly due to autologous proteolysis. This short half-life prevents the development of a robust immune response. Third, the catalytic domain sequence is highly conserved among species. Therefore, if elevated, the expected immune response must be weak. 6.2.1 Immunization Strategies In order to deal with these problems and increase the chance of success in obtaining MMP13 inhibitors combined with the catalytic bag, complex and sophisticated immunization strategies containing various formats of MMP13 were designed. Finally, the following immunization method was performed: (a) 3 llamas were immunized with a truncated MMP13 variant consisting of the catalytic domain and containing the mutation F72D that protects MMP13 from autologous proteolysis; (b) Three llamas were immunized with the same truncated MMP13 variant. The mutation system was the same as (a), but in addition to the mutation F72D, it also contained the mutation E120A that inactivates the enzyme function; (c) More than 3 llamas were immunized with the same truncated MMP13 variant. immunized with full-length proMMP13 protein; and (d) 3 llamas were immunized with a plasmid mixture encoding either a secreted proMMP13 variant (V123A) or a GPI-anchored proMMP13 variant (V123A). The V123A mutation described for MMP13 stimulates a weak interaction between Cys104 and the catalytic zinc ion, resulting in spontaneous self-activation. 6.2.2 Serum titration Determine serum titer against proMMP13 and catalytic domain (F72D). Animals immunized with the protein proMMP13 (c) or the DNA encoding proMMP13 V123A (d) generally showed good immune responses to proMMP13 but only weak responses to the catalytic domain (F72D). Animals immunized with the catalytic domain (F72D) (a) or the inactive catalytic domain (F72D, E120A) (b) showed no or only a weak immune response to the catalytic domain. 6.2.3 Library Construction Despite the low serum titers for the catalytic domain, the inventors are confident that large-scale screening can identify inhibitory binders to the catalytic pocket. RNA is extracted from PBL (primary blood lymphocytes) and used as a template for RT-PCR to amplify nanobodies encoding gene fragments. These fragments were selected and cloned into the phagemid vector pAX212 to produce phagemids displaying nanobodies fused to His6- and FLAG3-tags. Phage systems were prepared and stored according to standard procedures (Phage Display of Peptides and Proteins: A Laboratory Manual 1^st Edition, Brian K. Kay, Jill Winter, John McCafferty, Academic Press, 1996). The average size of the 18 immune libraries finally obtained is about 5 * 10⁸ Individually selected strains. 6.3 Preliminary screening Phage display selection was carried out with 18 immune libraries and two synthetic nanobody libraries. The libraries were targeted to different combinations of human proMMP13, activated human, rat and dog MMP13, and the human MMP13 catalytic domain (F72D) using different antigen presentation formats and underwent two to four rounds of intensive selection. Individual clones from the selection output were screened for binding to human and rat MMP13 in an ELISA using periplasmic extracts from E. coli cells expressing the nanobody and were measured using a broad-spectrum MMP fluorescent Fluorescent peptide assays target inhibitory activity by increasing fluorescence upon hydrolysis of the peptide of the photopeptide substrate. Nanobodies that showed binding in the ELISA but no inhibitory activity in the fluorescent peptide assay were further screened in a collagen lysis assay. The collagen dissolution assay uses natural substrates instead of peptide substitutes. When collagen and MMP13 have a much larger interaction surface than fluorescent peptides, it is speculated that nanobodies can interfere with collagen degradation but not peptide degradation. However, the collagen lysis assay is incompatible with periplasmic extracts and requires the use of purified nanobodies. 6.3.1 Test Series 1 In the first selection test series, MMP13 was presented as directly coated antigen. This resulted in high hit rates in human and rat MMP13 ELISAs (binding assays), including good diversity of nanobody binding to human MMP13, with a wide range of binding signals in the ELISA. Most nanobodies were found to be cross-reactive to rat MMP13. However, extremely low hit rates were obtained in the fluorescent peptide assay (inhibition assay). Furthermore, incomplete inhibition was observed and some of these Nanobodies showed no rat cross-reactivity. Since no monovalent fully inhibitory Nanobodies were obtained, the inventors speculate that the correct epitope was not targeted and the conditions used during selection were not optimal. However, test selection conditions were hampered in the absence of positive controls. It was eventually discovered that direct coating would interfere with enzyme activity. 6.3.2 Test series 2 In test series 2, the selection was performed with biotinylated MMP13 in solution. Using activated human and rat MMP13 captured via replacement of directly coated proMMP13 with unneutralized antibodies, however the ELISA hit rate was much lower than in trial series 1. In addition, the hit rate in the fluorescent peptide assay is also very low. However, a nanobody was found that completely inhibited the collagenolysis assay. Because the proform of the enzyme is used for binder enhancement, it is hypothesized that important epitopes are obscured by the propeptide. 6.3.3 Experiment Series 3 In view of the disappointing results of Experiment Series 1 and 2, the inventors chose to optimize the presentation of important epitopes in the catalytic domain by using activated MMP13 captured with unneutralizing antibodies. Also used in ELISA in the same capture format. This resulted in higher hit rates for both human and rat MMP13 in the ELISA. However, although the hit rate in the fluorescent peptide assay was slightly higher than in previous experimental series, the nanobodies still gave only incomplete inhibition and showed weak or no rat cross-reactivity, indicating that captured MMP13 The rendering is still sub-optimal. Although the three selected strains including C0101040E09 ("40E09") gave incomplete inhibition in the fluorescent peptide assay, they were found to be positive in the collagen dissolution assay. Family members of the family selection and sequence of 40E09: C0101PMP040E08, C0101pMP042A04, C0101pMP040B05, C0101pMP042D12, C0101pMP042A03, C0101PMP024A08 and C0101PMP040D01 (Reference Table A-1 and A-2). Sequence variability of the CDR regions is described in Tables 6.3.3A, 6.3.3B and 6.3.3C below. The amino acid sequence of the CDR of the selected strain 40E09 was used as a reference for comparison with the CDRs of family members (CDR1 starts at Kabat position 26, CDR2 starts at Kabat position 50, and CDR3 starts at Kabat position 95 ). 6.3.4 Trial Series 4 In the fourth trial series, in addition to the immune nanobody phage display libraries derived from immunization strategies (a) and (c), the phage display libraries derived from selection strategies (b) and (d) were also explored. library (refer to Example 6.2.1). The selection strategy focused on the MMP13 catalytic domain (F72D), which was used in solution. Hit rates in fluorescent peptide assays increased across libraries and many nanobodies showed complete inhibition. Inhibitory Nanobodies showed only poor binding in the ELISA, confirming that the captured MMP13 used in Experiment Series 3 presented suboptimal access to important epitopes. Therefore, a rat fluorescent peptide assay was used instead of ELISA to assess rat cross-reactivity. Representatives of families of nanobodies with confirmed inhibitory potential all have rat cross-reactivity, indicating that the epitopes recognized by this particular set of strains are located within the conserved MMP13 catalytic pocket. In summary, any conventional manipulation of MMP13 was found to interfere with enzymatic activity and TIMP-2 binding (data not shown). After changing and evaluating various parameters, including different antigens (such as proMMP13, catalytic domain (F72D), activated human MMP13, activated rat MMP13, activated dog MMP13), different assays (including ELISA, human fluorescent peptide assay, rat fluorescent peptide assay, and human collagen solubilization assay), different assay settings (including changes in coating conditions, in solution, and capturing MMP13) were found to be useful in identifying fully inhibitory nanoparticles The only successful strategies for antibodies were the use of the catalytic domain (F72D) or the selection of activated MMP13 species (Trial Series 4). 6.3.5 Pilot Panel Sequencing of inhibitory Nanobodies identified in fluorescent peptide or collagen lysis assays. Nanobodies can be classified into various families based on sequence information. Four families derived from test series 2 and 3 showed complete inhibition in the collagenolytic assay but no activity in the fluorescent peptide assay; additionally represented herein as "Profile 1" clones (ref. 40E09 and family members). Ten families derived from Screening Test Series 4 showed inhibitory activity in both the collagen dissolution assay and the fluorescent peptide assay; additionally represented herein as "Profile 2" clones. Select representative clones of each nanobody family, with a total of 14 representatives. The sequences of representative clones are described in Table A-1. 6.4 In vitro characterization of monovalent lead panels To further functionally characterize representative Nanobody clones, they were re-cloned into pAX129 according to standard procedures, transformed into E. coli and expressed and purified ( For example, Maussang et al. 2013 J Biol Chem 288(41): 29562-72). The selected strains then undergo various functional in vitro assays. 6.4.1 Enzymatic Assays The potency/efficacy of the nanobodies was tested in a fluorescent peptide assay targeting different MMP13 orthologs and in a human collagen lysis assay (both assays were also used during the screening , see Example 6.3). Additionally, a second collagen-based assay (fluorescent collagen assay) using higher collagen concentrations compared to the collagen lysis assay was developed to simulate high collagen concentration conditions in cartilage, anticipating MMP13 Inhibitors are active in it. In this assay, the intact FITC-labeled collagen substrate has low fluorescence due to the cross-quenching effect of the fluorophores. Upon cutting, quenching is lost and fluorescence increases. A summary of effectiveness in enzymatic assays is given in Table 6.4.1.In Figure 1, a dose response curve (fluorescent collagen assay) is depicted for the inhibitory effect of activated human MMP13 when high concentrations of bovine collagen I are used.Although Profile 1 nanobodies showed full efficacy in collagen lysis assays at low collagen concentrations, their efficacy in fluorescent collagen assays decreased at high collagen concentrations (Figure 1, left picture). The comparative drug MSC2392891A also showed weak efficacy in the fluorescent collagen assay (Figure 1). Overview 2 Representative nanobodies are effective and fully functional in both collagen dissolution assay and fluorescent collagen assay. Most nanobodies were more active than the comparator drugs in this assay. These Profile 2 representatives showed comparable potency (see Table 6.4.1) and efficacy (Figure 1, right panel) in human, macaque, rat, dog and bovine luciferase peptide assays. 6.4.2 Binding Assay Use an ELISA setup to assess binding affinity. However, this assay was found to be suitable only for assessing the affinity of Profile 1 Nanobodies, but not for Profile 2 Nanobodies (see Example 6.3). Describe the results in Table 6.4.2A.Overall, the Profile 1 Nanobodies had comparable binding affinities (i.e., less than 10-fold difference) for the three species tested. Although selection strain 40E09 had only the second-best binding affinity for human MMP13, it showed the best affinity across species. Because it was found that the profile 2 nanoantibody system competes with TIMP-2 for binding to MMP13, a TIMP-2 competitive ELISA was established and used to evaluate the affinity of the profile 2 nanoantibody. It is noteworthy that Profile 1 nanobodies do not compete with TIMP-2. Describe the results in Table 6.4.2B.Overall, the evaluation was similar to the potency obtained in the enzymatic assay, with Nanobodies 516G08, 529C12 and 62C02 showing the best inhibitory potency, and Nanobodies 59F06, 525C04 and 63F01 being the next best. 6.4.3 Selectivity assay In order to determine the selectivity of nanobodies for MMP13 over MMP1 and MMP14, a fluorescent peptide assay was used. MMP1 and MMP14 are two closely related members of the MMP family. Because Profile 1 Nanobodies did not show inhibition in the MMP13 fluorescent peptide assay, only Profile 2 Nanobodies may be tested. TIMP-2 (a non-selective MMP inhibitor) was used as a positive control in these assays. The results are depicted in Figure 2.All nanobodies are highly selective. They showed no MMP1 inhibitory effect and only observed very minor inhibition for some Nanobodies at high concentrations of MMP14. 6.4.4 Binning For antigen-binding, panels of monovalent Nanobodies derived from Profile 1 and Profile 2 were tested in a competitive ELISA against purified Nanobody 40E09 (Profile 1). The results of the competitive ELISA are shown in Figure 3.Overview 1 Nanobodies 32B08, 43B05, 43E10 and 40E09 (positive controls) all compete with 40E09. On the other hand, profile 2 nanobodies 59F06, 62C02, 63F01, 513C04, 516G08 and 517A01, and the negative control cAblys did not compete with 40E09. Therefore, the epitopic region bin of profile 1 nanobody (tentatively called "bin 1") seems to belong to a different bin from the epitope bin of profile 2 nanobody (tentatively called "bin 2"), which will also Reflects immunity and selection strategies. 6.5 Bivalent construct As demonstrated in Example 6.4.1 above, Profile 1 Nanobodies showed no inhibitory effect in the fluorescent peptide assay (refer to Table 6.4.1). The inventors set out to study the combined effects of Profile 1 Nanobodies and Profile 2 Nanobodies. As the best species cross-reactive binding agent (refer to Example 6.4.2), Nanobody 40E09 was selected as the profile 1 Nanobody representative. Overview 2, select three nanobodies: 516G08, 62C02 and 517A01. Profile 1 and Profile 2 nanoantibody systems are combined with the 35GS linker in a bivalent format (see Table 6.5; Nb(A)-35GS-Nb(B)). Bivalent nanobodies were selected and cloned into pAX205 according to standard procedures and transformed into methanophilic yeast (P. Pastoris ), expressed and purified. A rat fluorescent peptide assay was used to assess the potency of these bivalent constructs, but a lower MMP13 concentration (0.15 nM instead of 1.33 nM) was used compared to the screening setting to improve the sensitivity of the assay. The bivalent construct system was tested in this adapted fluorescent peptide assay as well as in a human fluorescent collagen assay. The obtained data are summarized in Table 6.5.The results indicate that a bivalent construct composed of a combination of Profile 1 Nanobodies and Profile 2 Nanobodies is more potent than those monovalent building blocks in an adapted rat fluorescent peptide assay, with up to 40-fold greater potency Improvement. Improved efficacy (up to 4-fold) was also observed in human fluorescent collagen assays. In both assays, the bivalent construct was equally potent as the positive, non-selective control TIMP-2. Thus, although Profile 1 Nanobodies are not particularly inhibitory by themselves, they improve the potency of Profile 2 Nanobodies when they are combined in bivalent constructs. 6.6 Biophysical Characterization To promote patient convenience, therapeutic compounds with low dosing frequency and high retention are preferred. Therefore, it is preferred that Nanobodies have high stability. To test stability, five representative profile 2 nanobodies and one representative profile 1 nanobody were subjected to biophysical characterization analysis. 6.6.1 Thermal Shift Assay The thermal stability of the wild-type anti-MMP13 Nanobody was studied in the Thermal Shift Assay (TSA). The results are described in Table 6.6.1.Tm values at pH 7 ranged from 65°C to 83°C, indicating good to very good stability characteristics. 6.6.2 Analytical SEC Polymerization and aggregation trends of a selected panel of 5 representative anti-MMP13 nanobodies were studied by analytical size exclusion chromatography (aSEC). The summarized results are shown in Table 6.6.2.Five representative nanobodies have retention times within the expected range of monovalent nanobodies (7.6 to 8.2 minutes), and for all nanobodies, the relative area of the main peak and the total recovery rate are both greater than 90%. Based on the biophysical properties of TSA and aSEC, all tested nanobodies were considered suitable for further development. 6.7 Selection of clones for further development and sequence optimization Based on the functional and biophysical characteristics of representative Nanobodies and bivalent constructs, 4 exemplary lead Nanobodies were selected for further development: 62C02, 529C12, 80A01 and bivalent construct C01010080 ("0080", consisting of Profile 2 Nanobody 517A01 and Profile 1 Nanobody 40E09). The inventors set out to optimize the amino acid sequence of the leader panel ("sequence optimization" or "SO"). In a sequence optimization approach, attempts are made to (1) eliminate sites for post-translational modifications (PTMs); (2) humanize the parent nanobody; and (3) eliminate sites for possible pre-existing antibodies. Antigenic determining region. At the same time, the functional and biophysical characteristics of the nanobody should preferably be maintained or even improved. 6.7.1 Post-translational modifications (PTMs) The post-translational modifications (PTMs) assessed were: Met-oxidation, Asn-deamidation, Asp-isomerization, Asn-glycation and pyroglutamic acid formation. Although the E1D mutation (which is typically incorporated to prevent pyroglutamic acid formation) was not analyzed during sequence optimization, it was included in the formatted Nanobodies. For all MMP13 lead nanobodies that underwent mutation, except 62C02, a 12-fold decrease in potency was observed. Therefore, it was decided not to incorporate the E1D mutation into the 62C02 building block. To assess possible PTM, the lead team was subjected to forced oxidation and temperature stress. No modifications were observed in the lead panel except C0101517A01 ("517A01") and C0101080A01 ("80A01"). Under forced oxidation and temperature stress conditions, C0101080A01 tends to Asp-isomerization and Met-oxidation. However, the degree of isomerization and oxidation differs under the conditions evaluated and is in each case just below or above the applied threshold values. Finally, amino acid residues 54-55, 100d-100e, M100j, and 101-102 were identified as the residues responsible for PTM. Notably, all of these residues are located in the CDR2 or CDR3 regions and therefore may be involved in target binding. To try to meet different needs, NKK libraries were constructed in which these residues were mutated and subsequently screened in a fluorescent peptide assay to assess possible loss of potency and to screen for biophysical properties (Tm). It was unexpectedly found that various positions can be mutated in these CDRs without any significant loss of potency, i.e. retention of over 80% inhibition. ¡ Regarding the D100dX library, 9 amino acid ("AA") substitutions showed >80% inhibition (E, G, A, P, T, R, M, W, and Y). ¡ Regarding the M100jX library, 16 AA substitutions showed >85% inhibition (all, except T, C, and H). ¡ Regarding the D101X library, 16 AA substitutions showed >90% inhibition (all, except D, F, and P). ¡ Regarding the Y102X library, most selected strains showed >90% inhibition. It is believed that amino acid position 102 may be mutated to any residue. The following conservative mutations are particularly good: D100dE and M100jL. A summary of preferred mutations in CDR regions is provided below in Tables 6.7.1A, 6.7.1B and 6.7.1C. The amino acid sequence of the CDR of the selected strain 80A01 was used as a reference for comparison with the CDRs of family members. CDR1 begins at amino acid residue 26 according to Kabat numbering, CDR2 begins at amino acid residue 50, and CDR3 begins at amino acid residue 95. X1=E,G,A,P,T,R,M,W,Y X2=A,R,N,D,E,Q,Z,G,I,L,K,F,P,S,W , Y and V X3=A, R, N, C, E, Q, Z, G, H, I, L, K, M, S, T, W, Y and V , C, E, Q, Z, G, H, I, L, K, M, F, P, S, T, W and V Under forced oxidation and temperature stress conditions, C0101517A01 tends to be deamidated. Finally, amino acid residues N100b and N101 were identified as residues sensitive to deamidation through temperature stress experiments. However, these residues are located in the CDR3 region and may be involved in target binding. Therefore, eliminating the cleidation site may have an impact on binding. In this case also an NKK library was constructed in which residues favoring deamidation were mutated and subsequently screened in a fluorescent peptide assay to assess possible loss of potency and to screen for biophysical properties (Tm). Completely unexpectedly, it was shown that mutation of N100b to Q or S or N101 to Q or V abrogates deamidation, but at the same time results in an increase in Tm of 1 to 3°C compared to the parent nanobody C0101517A01, whereas potency Comparable. The four preferred variants are shown in Table 6.7.1D, which also summarizes the results of the TSA and fluorescent peptide assays. A summary of preferred mutations in CDRs is described in the table below.A summary of preferred mutations in the CDR regions is provided below in Tables 6.7.1E, 6.7.1F and 6.7.1G. The amino acid sequence of the CDR of the selected strain 517A01 was used as a reference for comparison with the CDRs of other selected strains. According to Kabat numbering, CDR1 starts at amino acid residue 26, CDR2 starts at amino acid residue 50, and CDR3 starts at amino acid residue 95. 6.7.2 Humanization For humanization, the nanobody sequence is made more homologous to the human IGHV3-IGHJ germline consensus sequence. In addition to the nanobody "flag" residues, specific amino acids in the different framework regions between the nanobody and the human IGHV3-IGHJ germline consensus sequence are used to keep the protein structure, activity and stability intact. Change into human counterpart. 6.7.3 Pre-existing antibodies and anti-drug antibodies The inventors conducted an early risk assessment of clinical outcomes related to immunogenicity that led to sequence optimization strategies. This assessment includes the immunogenic potential of the drug candidate and the likelihood that anti-drug antibodies will impact both based on the mode of action and the nature of the final biotherapeutic molecule. The sequence of the Nanobody is evaluated against this to potentially (i) minimize the binding of any naturally occurring pre-existing antibodies, and (ii) reduce the likelihood of causing a therapeutically emergent immunogenic response. Mutations L11V and V89L were introduced into all MMP13 nanobodies. 6.7.4 Preferred SO-selected strains In Tables A-1 and A-2, the sequences are optimally described based on the sequence of the leading group, in which the sequences are based on possible pre-existing antibodies and anti-drug antibody PTMs, human source and epitope regions are elaborated. 6.8 Bispecific constructs Anti-MMP13 nanobodies should preferentially target sites of concern (such as joints) in order to inhibit the cartilage-degrading function of MMP13. Aggrecan is abundantly present in joints and is the major proteoglycan in the extracellular matrix (ECM) accounting for approximately 50% of the total protein content. Therefore, at least in theory, anchoring anti-MMP13 Nanobodies to aggrecan-binding agents could allow directing anti-MMP13 Nanobodies to relevant tissues and improve their retention. Set out to combine the anti-MMP13 binding agents of the invention with these aggrecan binding agents and test the efficacy of the resulting bispecific constructs in human fluorescent peptide assays and competitive ELISA assays. Various bispecific constructs containing aggrecan binders and MMP13 inhibitors were generated and tested as indicated in the table. The format and validity of typical results are described in Table 6.8A and Table 6.8B. The results shown in Tables 6.8A and 6.8B demonstrate that the combination of aggrecan binding agents and MMP13 inhibitors has no negative effect on the efficacy of the MMP13 inhibitors. Notably, in most cases, the lead group MMP13 inhibitors were as potent as the non-selective MMP inhibitor TIMP-2. Example 6.9 In vitro rat MMT model DMOAD study In order to further demonstrate the in vivo efficacy of the fusion of the MMP13 inhibitor and the CAP binding agent of the present invention, the surgically induced medial meniscal laceration (MMT) model in rats was used. Briefly, anti-MMP13 Nanobodies were coupled to CAP binding agent ("754" or C010100754). Operate on one knee of rats to induce OA-like symptoms. Treatment begins with IA injection 3 days after surgery. Histopathological examination was performed on day 42 after surgery. Interim and terminal serum samples were collected for exploratory biomarker analysis. Medial and total parenchymal cartilage degeneration widths were determined, as well as the percent reduction in cartilage degeneration. Twenty animals were used in each group. Suppression of cartilage degeneration in the medial tibia is shown in Figure 4. Results demonstrated that the MMP13-CAP construct significantly reduced cartilage width after 42 days compared to vehicle. These results indicate that (a) the CAP portion has no negative impact on the activity of the anti-MMP13 Nanobody (754), consistent with the results of Example 6.8; (b) the CAP portion is able to retain the anti-MMP13 Nanobody; and (c) the anti-MMP13 Nanobodies have a positive effect on cartilage width.The complete contents of all references cited throughout this application (including literature references, issued patents, published patent applications and patent applications also pending) are hereby expressly incorporated by reference. Incorporated into this article, specifically for the guidance mentioned above.

Figure 1: Dose-response curves of Profile 1 Nanobodies (left) and Profile 2 Nanobodies (right) in fluorescent collagen assays. Figure 2: Selectivity of MMP13 leader nanobodies. Figure 3: Competitive ELISA with 0.6 nM biotinylated 40E09 against the profile 1 and profile 2 nanobody panels on human full-length MMP13 coated with mouse anti-human MMP13 mAb (R&D Systems #MAB511). MMP13 was activated by incubation with APMA at 37°C for 90 minutes. Figure 4: Inhibition of cartilage degeneration using nanobodies in rat MMT model.

<110> Merck Patent GmbH Belgian company Ablynx N.V.

<120> Immunoglobulin binding to MMP13

<130> MEPAT.0104

<140> TW 107119216

<141> 2018-06-04

<150> EP17174402.2

<151> 2017-06-02

<160> 197

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<400> 103

<210> 104

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 104

<210> 105

<211> 2415

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 105

<210> 106

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 106

<210> 107

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 107

<210> 108

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 108

<210> 109

<211> 123

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 109

<210> 110

<211> 129

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 110

<210> 111

<211> 121

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 111

<210> 112

<211> 128

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 112

<210> 113

<211> 39

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 113

<210> 114

<211> 39

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 114

<210> 115

<211> 471

<212> PRT

<213> Homo sapiens

<220>

<223> Amino acid sequence

<400> 115

<210> 116

<211> 471

<212> PRT

<213> Chimpanzee

<220>

<223> Amino acid sequence

<400> 116

<210> 117

<211> 471

<212> PRT

<213> Rhesus macaque

<220>

<223> Amino acid sequence

<400> 117

<210> 118

<211> 496

<212> PRT

<213> Wolf

<220>

<223> Amino acid sequence

<400> 118

<210> 119

<211> 471

<212> PRT

<213> Cattle

<220>

<223> Artificial sequence

<400> 119

<210> 120

<211> 472

<212> PRT

<213> mice

<220>

<223> Amino acid sequence

<400> 120

<210> 121

<211> 472

<212> PRT

<213> Brown Rat

<220>

<223> Amino acid sequence

<400> 121

<210> 122

<211> 471

<212> PRT

<213> Chicken

<220>

<223> Amino acid sequence

<400> 122

<210> 123

<211> 23

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 123

<210> 124

<211> 34

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 124

<210> 125

<400> 125

000

<210> 126

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 126

<210> 127

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 127

<210> 128

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 128

<210> 129

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 129

<210> 130

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 130

<210> 131

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 131

<210> 132

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 132

<210> 133

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 133

<210> 134

<211> 25

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 134

<210> 135

<211> 30

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 135

<210> 136

<211> 35

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 136

<210> 137

<211> 40

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 137

<210> 138

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 138

<210> 139

<211> 24

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 139

<210> 140

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 140

<210> 141

<211> 62

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 141

<210> 142

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 142

<210> 143

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 143

<210> 144

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 144

<210> 145

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 145

<210> 146

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 146

<210> 147

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 147

<210> 148

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 148

<210> 149

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 149

<210> 150

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 150

<210> 151

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 151

<210> 152

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 152

<210> 153

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 153

<210> 154

<211> 118

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 154

<210> 155

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 155

<210> 156

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 156

<210> 157

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 157

<210> 158

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 158

<210> 159

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 159

<210> 160

<211> 278

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 160

<210> 161

<211> 269

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 161

<210> 162

<211> 270

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 162

<210> 163

<211> 278

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 163

<210> 164

<211> 269

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 164

<210> 165

<211> 270

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 165

<210> 166

<211> 121

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 166

<210> 167

<211> 122

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 167

<210> 168

<211> 125

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 168

<210> 169

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 169

<210> 170

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 170

<210> 171

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 171

<210> 172

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 172

<210> 173

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 173

<210> 174

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 174

<210> 175

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 175

<210> 176

<211> 277

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 176

<210> 177

<211> 433

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 177

<210> 178

<211> 280

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 178

<210> 179

<211> 284

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 179

<210> 180

<211> 440

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 180

<210> 181

<211> 287

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 181

<210> 182

<211> 438

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 182

<210> 183

<211> 434

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 183

<210> 184

<211> 590

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 184

<210> 185

<211> 437

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 185

<210> 186

<211> 278

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 186

<210> 187

<211> 278

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 187

<210> 188

<211> 283

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 188

<210> 189

<211> 279

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 189

<210> 190

<211> 435

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 190

<210> 191

<211> 282

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 191

<210> 192

<211> 434

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 192

<210> 193

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 193

<210> 194

<211> 155

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 194

<210> 195

<211> 162

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 195

<210> 196

<211> 312

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 196

<210> 197

<211> 157

<212> PRT

<213> Artificial sequence

<220>

<223> Amino acid sequence

<400> 197

Claims (41)

A polypeptide comprising at least 1 immunoglobulin single variable domain (ISVD) that binds to a matrix metalloproteinase (MMP), wherein the MMP is MMP13, and wherein the ISVD that binds to MMP13 includes 3 complementarity determining regions (respectively are CDR1 to CDR3), wherein the ISVD includes a combination of CDR1, CDR2 and CDR3 selected from the group consisting of: - CDR1 is SEQ ID NO: 27, CDR2 is SEQ ID NO: 42, and CDR3 is SEQ ID NO: 56; - CDR1 is SEQ ID NO: 28, CDR2 is SEQ ID NO: 43, and CDR3 is SEQ ID NO: 107; - CDR1 is SEQ ID NO: 28, CDR2 is SEQ ID NO: 43, and CDR3 is SEQ ID NO. NO: 57; - CDR1 is SEQ ID NO: 25, CDR2 is SEQ ID NO: 40, and CDR3 is SEQ ID NO: 54; - CDR1 is SEQ ID NO: 26, CDR2 is SEQ ID NO: 41, and CDR3 is SEQ ID NO: 106; - CDR1 is SEQ ID NO: 26, CDR2 is SEQ ID NO: 41, and CDR3 is SEQ ID NO: 55; and - CDR1 is SEQ ID NO: 23, CDR2 is SEQ ID NO: 37, and CDR3 is SEQ ID NO: 52. According to the polypeptide of claim 1, the ISVD that binds to MMP13 does not bind to MMP1 or MMP14 (membrane type). According to the polypeptide of claim 1 or 2, the ISVD is composed of or essentially consists of 4 framework regions (FR1 to FR4 respectively) and the 3 complementarity determining regions CDR1, CDR2 and CDR3. The polypeptide according to claim 1 or 2, wherein the polypeptide is SEQ ID NO: 1 or a polypeptide having at least 95% sequence identity with SEQ ID NO: 1. According to the polypeptide of claim 1 or 2, CDR1 is SEQ ID NO: 27, CDR2 is SEQ ID NO: 42, and CDR3 is SEQ ID NO: 56. According to the polypeptide of claim 1 or 2, the ISVD is selected from the group consisting of: SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10 and 1. The polypeptide according to claim 1 or 2, wherein the polypeptide binds to MMP13 with a K _D between 1E ^-07 M and 1E ^-13 M. The polypeptide according to claim 7, wherein the polypeptide binds to MMP13 with a _KD selected from the group consisting of: at most 1E ^-07 M, less than 1E ^-08 M or 1E ^-09 M, or even Below 1E ^-10 M. The polypeptide according to claim 1 or 2, wherein the polypeptide inhibits the activity of MMP13 with an IC ₅₀ between 1E ^-07 M and 1E ^-12 M. The polypeptide according to claim 9 of the patent application, wherein the polypeptide inhibits the activity of MMP13 with an IC ₅₀ of at most 1E ^-07 M. The polypeptide according to item 10 of the patent application, wherein the polypeptide inhibits MMP13 with an IC ₅₀ of 1E ^-08 M, 5E ^-09 M, or 4E ^-9 M, 3E ^-9 M, 2E ^-9 M, or 1E ^-9 M of activity. The polypeptide according to claim 1 or 2, wherein the polypeptide binds to MMP13 with an EC ₅₀ between 1E ^-07 M and 1E ^-12 M. The polypeptide according to claim 1 or 2, wherein the polypeptide binds to MMP13 with an off-rate lower than 5E ^-04 s ^-1 . According to the polypeptide of item 1 or 2 of the patent application, the MMP13 is human MMP13, rat MMP13, dog MMP13, bovine MMP13, or crab-eating macaque MMP13. According to the polypeptide of claim 14, the human MMP13 is SEQ ID NO: 115. A polypeptide according to item 1 or 2 of the patent application, wherein the polypeptide antagonizes The activity of MMP13, wherein the activity is selected from: (i) protease activity; and (ii) binding of collagen to a thrombin-like domain. The polypeptide according to claim 16, wherein the polypeptide blocks at least 20% of the binding of MMP13 to collagen and/or aggrecan. The polypeptide according to claim 1 or 2, wherein the polypeptide inhibits the protease activity of MMP13. According to the polypeptide of claim 18 of the patent application, the protease activity is proteolysis of type II collagen. The polypeptide according to item 1 or 2 of the patent application, which contains at least 2 ISVDs, wherein at least 1 ISVD specifically binds to MMP13, and wherein the polypeptide contains 2, 3 or 4 ISVDs. According to the polypeptide of claim 20, the at least two ISVDs specifically bind to MMP13. The polypeptide according to item 1 of the patent application, which contains 2, 3 or 4 ISVDs, each of which specifically binds to MMP13 individually, wherein a) at least the "first" ISVD is bound to the first antigenic determinant and antigen of MMP13 determines specific binding of a region, portion, domain, subunit or conformation; and wherein b) at least the "second" ISVD specifically binds to a second epitope, epitope, part, domain, subunit or configuration of MMP13 that is different from the first epitope, epitope, respectively. Region, part, domain, subunit or configuration. According to the polypeptide of claim 22, the "first" ISVD that specifically binds to MMP13 is selected from the group consisting of: SEQ ID NO: 111, 11, 110, 10, 112, 12, 109 and 9. According to the polypeptide of claim 22 or 23, the "second" ISVD that specifically binds to MMP13 is SEQ ID NO: 1. The polypeptide according to item 1 or 2 of the aforementioned patent application, further comprising one or two ISVDs that specifically bind to aggrecan, wherein the ISVDs that specifically bind to aggrecan can be the same or different, wherein The ISVD that specifically binds aggrecan is independently selected from: SEQ ID NO: 166 to 168. According to the polypeptide of claim 25, the ISVD that specifically binds to aggrecan specifically binds to human aggrecan [SEQ ID NO: 105]. According to the polypeptide of claim 25 of the patent application, which is polymerized with aggregin The ISVD system specifically binds sugars to dog aggrecan, bovine aggrecan, rat aggrecan, porcine aggrecan, mouse aggrecan, rabbit aggrecan, and crab-eating macaque. Specific binding to aggrecan and/or rhesus aggrecan. According to the polypeptide of claim 25, the ISVD specifically binding to aggrecan binds to cartilage tissue. The polypeptide according to claim 1 or 2, wherein the polypeptide has a stability of at least 7 days in synovial fluid (SF) at 37°C. The polypeptide according to item 1 or 2 of the patent application further includes an ISVD that binds to serum albumin, wherein the ISVD that binds to serum albumin basically consists of 4 framework regions (FR1 to FR4 respectively) and 3 complementary regions. Determining regions (respectively CDR1 to CDR3), wherein CDR1 is SEQ ID NO: 157, CDR2 is SEQ ID NO: 158, and CDR3 is SEQ ID NO: 159. According to the polypeptide of claim 30, the ISVD binding to serum albumin is selected from the group consisting of: ALB135 (SEQ ID NO: 193), ALB129 (SEQ ID NO: 144), ALB8 (SEQ ID NO:142), ALB23 (SEQ ID NO:143) and ALB132 (SEQ ID NO:145). The polypeptide according to claim 20, wherein the at least two ISVDs are directly linked to each other or linked via a linker. According to the polypeptide of claim 32, the linker is selected from the group consisting of: SEQ ID NO: 125 to 141. According to the polypeptide of item 32 of the patent application, the linker is SEQ ID NO: 129. The polypeptide according to claim 1 or 2, further comprising a C-terminal extension, wherein the C-terminal extension is C-terminal extension (X)n, where n is 1 to 10; and each X is independently selected An amino acid residue, wherein the amino acid residue is independently selected from the group consisting of: alanine (A), glycine (G), valine (V), leucine (L) ) and isoleucine(I). The polypeptide according to item 1 or 2 of the patent application, wherein the polypeptide is the same as SEQ ID NO: 1, 9, 10, 11, 12, 109, 110, 111, 112, 160, 161, 162, 163, 164, 165 Any one of , 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 or 192 has at least 80%, 90%, 95% or 100 % sequence identity. The use of a polypeptide according to any one of items 1 to 36 of the patent application in manufacturing a medicament for treating and preventing individual diseases or disorders, which involves MMP13 activity. The use according to item 37 of the patent application, wherein the disease or disease is selected from the group consisting of: joint lesions and chondrodystrophies, arthritic diseases, osteoarthritis, rheumatoid arthritis, gouty arthritis , psoriatic arthritis, traumatic rupture or detachment, achondroplasia, costochondritis, spondylosis, intervertebral disc herniation, lumbar disc degenerative disease, degenerative joint disease and relapsing polychondritis, osteochondral dissecans inflammation and aggrecanopath. The use of a polypeptide according to any one of items 1 to 36 of the patent application for manufacturing a medicament. Polypeptides according to item 1 or 2 of the patent application, which are used for the treatment or prevention of symptoms consisting of: joint lesions and chondrodystrophies, arthritic diseases, osteoarthritis, rheumatoid arthritis, gouty joints arthritis, psoriatic arthritis, traumatic rupture or detachment, achondroplasia, costochondritis, aplasia of the vertebrae, disc herniation, degenerative lumbar disc disease, degenerative joint disease and relapsing polychondritis, osteosarcoma Chondritis and aggrecan lesions. The polypeptide according to claim 1 or 2, wherein the polypeptide cross-blocks with at least one of the polypeptides according to any one of SEQ ID NO: 111, 11, 112, 12, 109, 9, 110, 10 and 1 The MMP13 binding and/ Or is cross-blocked from binding to MMP13 by at least one polypeptide according to any one of SEQ ID NOs: 111, 11, 112, 12, 109, 9, 110, 10 and 1.